LOCUS       pTwist_CMV_rbIgG        7605 bp    DNA     circular SYN 18-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7605)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 18, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7605
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1175..7605,1..1174)
                     /organism="pTwistCMVrIgG1 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..2146
                     /label=rbIgG1
     misc_feature    2171..2759
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2642..2653)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2812..2933
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      3048..3094
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3204..3275
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        3320..3516
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3367..3502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3530..4324
                     /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    4335..4559
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4755..5343)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6089..6193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6194..6262,6263..7054)
                     /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      7067..7203
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7348..7419
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    7477..7525
                     /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 ccgcggtcag cccaaggctc cgagcgtttt
     1201 cccattggca ccttgctgtg gggatactcc ttccagtact gtgaccctcg gttgcctggt
     1261 taagggctac ctgccggaac ctgtgaccgt tacttggaac tctggaaccc tcacgaatgg
     1321 ggtgcgcacc ttccctagtg tgcggcaaag ctctggcctg tacagcctgt ctagcgtggt
     1381 ctccgtcaca agctcttccc agccagtgac ctgcaacgtt gctcaccccg ccactaacac
     1441 caaggtcgac aagaccgtcg caccgagcac ttgtagcaaa cctacgtgtc ctccacccga
     1501 actgttggga ggcccctccg tgttcatctt tcctccaaaa ccaaaggaca ctctcatgat
     1561 ctccaggacc cccgaggtca cctgcgtcgt ggtcgatgtc tcccaggatg accccgaagt
     1621 ccagtttacc tggtatatca ataacgagca agtccgcact gcgcggcctc cgttgcgcga
     1681 gcaacagttc aacagtacta tcagagtggt ctctaccctg ccgatcactc atcaagactg
     1741 gctgcggggt aaggagttca agtgcaaggt ccataacaaa gcgcttccag ctcccatcga
     1801 gaaaacaatc tccaaggctc gcggccaacc tctggaacca aaggtgtaca ccatggggcc
     1861 ccctcgcgag gaactcagct cccgtagtgt ttcccttaca tgcatgatca acggcttcta
     1921 tcctagcgac atttcagtag agtgggaaaa aaacgggaag gctgaggata actataaaac
     1981 tacccccgcc gtcctggatt ccgacggctc ctactttctg tataacaagc tgtctgtccc
     2041 caccagcgag tggcagcgcg gtgacgtatt cacgtgttcc gtgatgcacg aggcgctcca
     2101 caaccactat acacagaagt ccatttctcg ttccccgggc aagtgaatcg atttctggat
     2161 ccgcaggcct aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa
     2221 ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat
     2281 tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta
     2341 tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc
     2401 aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt
     2461 ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg
     2521 ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc
     2581 atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc
     2641 ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct
     2701 tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc
     2761 tgtctagctt gactgactga gatacagcgt accttcagct cacagacatg ataagataca
     2821 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2881 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2941 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaaagca
     3001 agtaaaacct ctacaaatgt ggtattggcc catctctatc ggtatcgtag cataacccct
     3061 tggggcctct aaacgggtct tgaggggttt tttgtgcccc tcgggccgga ttgctatcta
     3121 ccgaggcact ggtgtactga tagagccggg ctagatgtta actgtcagct atcctgcagg
     3181 cgatctctcg atttcgatca agacattcct ttaatggtct tttctggaca ccactagggg
     3241 tcagaagtag ttcatcaaac tttcttccct ccctaatctc attggttacc ttgggctatc
     3301 gaaacttaat taagcgatct gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     3361 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tcgctgacta
     3421 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     3481 tgaggaggct tttttggagg cctaggcttt tgcaaaggag gtagccaaca tgattgaaca
     3541 agatggattg cacgcaggtt ctcccgccgc ttgggtggag aggctattcg gctatgactg
     3601 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3661 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactcc aggacgaggc
     3721 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3781 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3841 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3901 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3961 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4021 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgg atgcccgacg gcgaggatct
     4081 cgtcgtgacc cacggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4141 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4201 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4261 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4321 ctagtatgta agccctgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     4381 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     4441 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     4501 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggt
     4561 taattaacca gtcaagtcag ctacttggcg agatcgactt gtctgggttt cgactacgct
     4621 cagaattgcg tcagtcaagt tcgatctggt ccttgctatt gcacccgttc tccgattacg
     4681 agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4741 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4801 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4861 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4921 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4981 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     5041 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     5101 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5161 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5221 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5281 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5341 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5401 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5461 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5521 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgca
     5581 tttaaatttc cgaactctcc aaggccctcg tcggaaaatc ttcaaacctt tcgtccgatc
     5641 catcttgcag gctacctctc gaacgaacta tcgcaagtct cttggccggc cttgcgcctt
     5701 ggctattgct tggcagcgcc tatcgccagg tattactcca atcccgaata tccgagatcg
     5761 ggatcacccg agagaagttc aacctacatc ctcaatcccg atctatccga gatccgagga
     5821 atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc ctctcagtgc gagtctcgac
     5881 gatccatatc gttgcttggc agtcagccag tcggaatcca gcttgggacc caggaagtcc
     5941 aatcgtcaga tattgtactc aagcctggtc acggcagcgt accgatctgt ttaaacctag
     6001 atattgatag tctgatcggt caacgtataa tcgagtccta gcttttgcaa acatctatca
     6061 agagacagga tcagcaggag gctttcgccg cggaacccct atttgtttat ttttctaaat
     6121 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     6181 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     6241 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     6301 tcagttgggt gcgcgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6361 gagttttcgc cccgaagaac gctttccaat gatgagcact tttaaagttc tgctatgtgg
     6421 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6481 tcagaatgac ttggttgagt attcaccagt cacagaaaag catcttacgg atggcatgac
     6541 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6601 tctgacaacg attggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6661 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6721 tgacaccacg atgcctgtag caatggcaac aaccttgcgt aaactattaa ctggcgaact
     6781 acttactcta gcttcccggc aacagttgat agactggatg gaggcggata aagttgcagg
     6841 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6901 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6961 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     7021 tgagataggt gcctcactga ttaagcattg gtaaccgatt ctaggtgcat tggcgcagaa
     7081 aaaaatgcct gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg
     7141 gatacggctt ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct
     7201 taagatcgtt taaactcgac tctggctcta tcgaatctcc gtcgtttcga gcttacgcga
     7261 acagccgtgg cgctcatttg ctcgtcgggc atcgaatctc gtcagctatc gtcagcttac
     7321 ctttttggca gcgatcgcgg ctcccgacat cttggaccat tagctccaca ggtatcttct
     7381 tccctctagt ggtcataaca gcagcttcag ctacctctca atttgcgtat agagcgggtc
     7441 atcgaagggt tatacactcg taatcgttgc gttacacaca caaaaaacca acacacatcc
     7501 atcttcgatg gatagcgatt ttattatcta actgctgatc gagtgtagcc agatctagta
     7561 atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
