LOCUS       pTwist_CMV_hIgG1        7626 bp    DNA     circular SYN 10-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7626)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 10, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1175..7626,1..1174)
                     /organism="pTwistCMVhIgG1(LALA+PG+YTE) in pTwist CMV BG 
                     WPRE Neo v2.0"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        306..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          642..1117
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     misc_feature    1174^1175
                     /label=Your gene here
     misc_feature    1175..2167
                     /label=hIgG1L234A_L235A+P329G+M252Y_S254T_T256E
     misc_feature    2192..2780
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2663..2674)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2833..2954
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      3069..3115
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3225..3296
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        3341..3537
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3388..3523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3551..4345
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4356..4580
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4776..5364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6110..6214
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6215..6283,6284..7075)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      7088..7224
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7369..7440
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    7498..7546
                     /note="synthetic polyadenylation signal"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catgctgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tcccctcgaa
      601 gcttacatgt ggtaccgagc tcggatcctg agaacttcag ggtgagtcta tgggaccctt
      661 gatgttttct ttccccttct tttctatggt taagttcatg tcataggaag gggagaagta
      721 acagggtaca catattgacc aaatcagggt aattttgcat ttgtaatttt aaaaaatgct
      781 ttcttctttt aatatacttt tttgtttatc ttatttctaa tactttccct aatctctttc
      841 tttcagggca ataatgatac aatgtatcat gcctctttgc accattctaa agaataacag
      901 tgataatttc tgggttaagg caatagcaat atttctgcat ataaatattt ctgcatataa
      961 attgtaactg atgtaagagg tttcatattg ctaatagcag ctacaatcca gctaccattc
     1021 tgcttttatt ttatggttgg gataaggctg gattattctg agtccaagct aggccctttt
     1081 gctaatcatg ttcatacctc ttatcttcct cccacagctc ctgggcaacg tgctggtctg
     1141 tgtgctggcc catcactttg gcaaaggcgg ccgcgctagc acaaagggcc ccagcgtgtt
     1201 ccctctggct cctagcagca agtctacaag cggaggaaca gccgctctgg gctgcctggt
     1261 caaggattac tttcccgagc ctgtgaccgt gtcctggaat tctggcgctc tgacaagcgg
     1321 cgtgcacacc tttccagctg tgctgcaaag ctccggcctg tactctctga gcagcgtggt
     1381 cacagtgcca agctctagcc tgggcaccca gacctacatc tgcaatgtga accacaagcc
     1441 tagcaacacc aaggtggaca agagagtgga acccaagagc tgcgacaaga cccacacctg
     1501 tcctccatgt cctgctccag aagctgctgg cggcccatcc gtgtttctgt tccctccaaa
     1561 gcctaaggac accctgtaca tcacccgcga gcctgaagtg acctgtgtgg tggtggatgt
     1621 gtcccacgag gacccagaag tgaagttcaa ttggtacgtg gacggcgtgg aagtgcacaa
     1681 cgccaagacc aagcctagag aggaacagta caacagcacc tacagagtgg tgtccgtgct
     1741 gaccgtgctg caccaggatt ggctgaacgg caaagagtac aagtgcaagg tgtccaacaa
     1801 ggccctgggc gctcccatcg agaaaaccat ctctaaggcc aagggccagc ctcgcgaacc
     1861 ccaggtttac acactgcctc caagccggga agagatgacc aagaaccagg tgtccctgac
     1921 ctgcctcgtg aagggcttct acccttccga tatcgccgtg gaatgggaga gcaatggcca
     1981 gccagagaac aactacaaga caacccctcc tgtgctggac agcgacggct cattcttcct
     2041 gtacagcaag ctgacagtgg acaagtccag atggcagcag ggcaacgtgt tcagctgcag
     2101 cgtgatgcac gaggccctgc acaaccacta cacccagaaa agcctgtctc tgagccccgg
     2161 caagtgaatc gatttctgga tccgcaggcc taatcaacct ctggattaca aaatttgtga
     2221 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2281 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2341 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2401 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2461 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2521 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2581 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2641 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2701 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     2761 cctttgggcc gcctccccgc ctgtctagct tgactgactg agatacagcg taccttcagc
     2821 tcacagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2881 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2941 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     3001 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtattggc ccatctctat
     3061 cggtatcgta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgtgccc
     3121 ctcgggccgg attgctatct accgaggcac tggtgtactg atagagccgg gctagatgtt
     3181 aactgtcagc tatcctgcag gcgatctctc gatttcgatc aagacattcc tttaatggtc
     3241 ttttctggac accactaggg gtcagaagta gttcatcaaa ctttcttccc tccctaatct
     3301 cattggttac cttgggctat cgaaacttaa ttaagcgatc tgcatctcaa ttagtcagca
     3361 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3421 tctccgcccc atcgctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3481 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagga
     3541 ggtagccaac atgattgaac aagatggatt gcacgcaggt tctcccgccg cttgggtgga
     3601 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     3661 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     3721 gaatgaactc caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     3781 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     3841 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     3901 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     3961 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4021 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4081 gatgcccgac ggcgaggatc tcgtcgtgac ccacggcgat gcctgcttgc cgaatatcat
     4141 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4201 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4261 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4321 tcgccttctt gacgagttct tctagtatgt aagccctgtg ccttctagtt gccagccatc
     4381 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4441 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4501 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4561 ggatgcggtg ggctctatgg ttaattaacc agtcaagtca gctacttggc gagatcgact
     4621 tgtctgggtt tcgactacgc tcagaattgc gtcagtcaag ttcgatctgg tccttgctat
     4681 tgcacccgtt ctccgattac gagtttcatt taaatcatgt gagcaaaagg ccagcaaaag
     4741 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4801 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4861 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4921 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4981 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5041 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5101 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5161 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5221 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5281 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5341 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5401 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5461 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5521 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5581 tttcgttcat ccatagttgc atttaaattt ccgaactctc caaggccctc gtcggaaaat
     5641 cttcaaacct ttcgtccgat ccatcttgca ggctacctct cgaacgaact atcgcaagtc
     5701 tcttggccgg ccttgcgcct tggctattgc ttggcagcgc ctatcgccag gtattactcc
     5761 aatcccgaat atccgagatc gggatcaccc gagagaagtt caacctacat cctcaatccc
     5821 gatctatccg agatccgagg aatatcgaaa tcggggcgcg cctggtgtac cgagaacgat
     5881 cctctcagtg cgagtctcga cgatccatat cgttgcttgg cagtcagcca gtcggaatcc
     5941 agcttgggac ccaggaagtc caatcgtcag atattgtact caagcctggt cacggcagcg
     6001 taccgatctg tttaaaccta gatattgata gtctgatcgg tcaacgtata atcgagtcct
     6061 agcttttgca aacatctatc aagagacagg atcagcagga ggctttcgcc gcggaacccc
     6121 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6181 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6241 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6301 gaaagtaaaa gatgctgaag atcagttggg tgcgcgagtg ggttacatcg aactggatct
     6361 caacagcggt aagatccttg agagttttcg ccccgaagaa cgctttccaa tgatgagcac
     6421 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6481 cggtcgccgc atacactatt ctcagaatga cttggttgag tattcaccag tcacagaaaa
     6541 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6601 taacactgcg gccaacttac ttctgacaac gattggagga ccgaaggagc taaccgcttt
     6661 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6721 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caaccttgcg
     6781 taaactatta actggcgaac tacttactct agcttcccgg caacagttga tagactggat
     6841 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6901 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6961 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     7021 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaaccgat
     7081 tctaggtgca ttggcgcaga aaaaaatgcc tgatgcgacg ctgcgcgtct tatactccca
     7141 catatgccag attcagcaac ggatacggct tccccaactt gcccacttcc atacgtgtcc
     7201 tccttaccag aaatttatcc ttaagatcgt ttaaactcga ctctggctct atcgaatctc
     7261 cgtcgtttcg agcttacgcg aacagccgtg gcgctcattt gctcgtcggg catcgaatct
     7321 cgtcagctat cgtcagctta cctttttggc agcgatcgcg gctcccgaca tcttggacca
     7381 ttagctccac aggtatcttc ttccctctag tggtcataac agcagcttca gctacctctc
     7441 aatttgcgta tagagcgggt catcgaaggg ttatacactc gtaatcgttg cgttacacac
     7501 acaaaaaacc aacacacatc catcttcgat ggatagcgat tttattatct aactgctgat
     7561 cgagtgtagc cagatctagt aatcaattac ggggtcatta gttcatagcc catatatgga
     7621 gttccg
//
