LOCUS       pTwist_CMV_bispe        7623 bp    DNA     circular SYN 04-FEB-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7623)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1175..7623,1..1174)
                     /organism="pTwistCMVbispecifichIgG1(hole) 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..2164
                     /label=bispecific hIgG1 full hole
     misc_feature    2189..2777
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2660..2671)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2830..2951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      3066..3112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3222..3293
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        3338..3534
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3385..3520
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3548..4342
                     /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    4353..4577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4773..5361)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6107..6211
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6212..6280,6281..7072)
                     /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      7085..7221
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7366..7437
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    7495..7543
                     /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 ccgcgcttcc accaagggcc cctctgtttt
     1201 cccattggct ccttcatcaa agtctacaag cggcggcacc gctgctctag gatgtctcgt
     1261 caaagactac ttccctgaac ctgttacagt gagctggaac agtggcgcgc taacgtctgg
     1321 agtgcacaca ttcccagctg tgctgcagag ttctggcctg tattctctgt ctagcgtggt
     1381 gacagtgcct tccagttccc tcggtacgca gacttacatt tgcaacgtaa accacaaacc
     1441 ctcaaacaca aaagtggata agaaggttga gcctaaatct tgcgataaga ctcacacctg
     1501 tccgccatgc cccgccccgg aacttctggg tggccctagt gtgtttctgt tccctcccaa
     1561 acctaaggac acactcatga ttagcagaac tccagaagtc acgtgcgtgg tcgtggatgt
     1621 gtcccacgag gacccagagg tgaagtttaa ctggtacgta gatggcgtgg aggttcacaa
     1681 cgcaaagact aagccgcgcg aagagcagta caactccaca tacagagtcg tgtctgtgct
     1741 caccgtgctg catcaggact ggctgaatgg caaagagtac aagtgcaaag tgtccaacaa
     1801 ggctctgccc gctccaatcg agaagacgat ctccaaggcc aagggtcagc cccgtgagcc
     1861 ccaggtgtat accctgccgc cctctcgtga ggaaatgacc aagaaccagg tgagcctgag
     1921 ttgtgcagtg aagggcttct atccttccga tatcgccgtc gagtgggaaa gcaacgggca
     1981 gccagagaat aactacaaga ccacaccccc tgtgctcgac tccgacgggt catttttcct
     2041 ggtatctaaa ctcaccgttg acaagagcag gtggcagcaa ggaaacgtgt tctcctgctc
     2101 cgttatgcac gaggccctgc ataaccacta cacgcagaaa tccctcagcc tgagccccgg
     2161 gtgaatcgat ttctggatcc gcaggcctaa tcaacctctg gattacaaaa tttgtgaaag
     2221 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2281 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2341 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2401 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2461 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2521 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2581 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     2641 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     2701 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     2761 ttgggccgcc tccccgcctg tctagcttga ctgactgaga tacagcgtac cttcagctca
     2821 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     2881 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     2941 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     3001 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tattggccca tctctatcgg
     3061 tatcgtagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgtgcccctc
     3121 gggccggatt gctatctacc gaggcactgg tgtactgata gagccgggct agatgttaac
     3181 tgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     3241 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     3301 tggttacctt gggctatcga aacttaatta agcgatctgc atctcaatta gtcagcaacc
     3361 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3421 ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3481 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaggaggt
     3541 agccaacatg attgaacaag atggattgca cgcaggttct cccgccgctt gggtggagag
     3601 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3661 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3721 tgaactccag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3781 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3841 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3901 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3961 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4021 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat
     4081 gcccgacggc gaggatctcg tcgtgaccca cggcgatgcc tgcttgccga atatcatggt
     4141 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4201 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4261 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4321 ccttcttgac gagttcttct agtatgtaag ccctgtgcct tctagttgcc agccatctgt
     4381 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     4441 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     4501 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     4561 tgcggtgggc tctatggtta attaaccagt caagtcagct acttggcgag atcgacttgt
     4621 ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc ttgctattgc
     4681 acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca gcaaaaggcc
     4741 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4801 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4861 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4921 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4981 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5041 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5101 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5161 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5221 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5281 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5341 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5401 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5461 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5521 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5581 cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc ggaaaatctt
     5641 caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc gcaagtctct
     5701 tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta ttactccaat
     5761 cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct caatcccgat
     5821 ctatccgaga tccgaggaat atcgaaatcg gggcgcgcct ggtgtaccga gaacgatcct
     5881 ctcagtgcga gtctcgacga tccatatcgt tgcttggcag tcagccagtc ggaatccagc
     5941 ttgggaccca ggaagtccaa tcgtcagata ttgtactcaa gcctggtcac ggcagcgtac
     6001 cgatctgttt aaacctagat attgatagtc tgatcggtca acgtataatc gagtcctagc
     6061 ttttgcaaac atctatcaag agacaggatc agcaggaggc tttcgccgcg gaacccctat
     6121 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6181 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6241 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6301 agtaaaagat gctgaagatc agttgggtgc gcgagtgggt tacatcgaac tggatctcaa
     6361 cagcggtaag atccttgaga gttttcgccc cgaagaacgc tttccaatga tgagcacttt
     6421 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6481 tcgccgcata cactattctc agaatgactt ggttgagtat tcaccagtca cagaaaagca
     6541 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6601 cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa ccgctttttt
     6661 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6721 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa ccttgcgtaa
     6781 actattaact ggcgaactac ttactctagc ttcccggcaa cagttgatag actggatgga
     6841 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6901 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6961 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7021 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aaccgattct
     7081 aggtgcattg gcgcagaaaa aaatgcctga tgcgacgctg cgcgtcttat actcccacat
     7141 atgccagatt cagcaacgga tacggcttcc ccaacttgcc cacttccata cgtgtcctcc
     7201 ttaccagaaa tttatcctta agatcgttta aactcgactc tggctctatc gaatctccgt
     7261 cgtttcgagc ttacgcgaac agccgtggcg ctcatttgct cgtcgggcat cgaatctcgt
     7321 cagctatcgt cagcttacct ttttggcagc gatcgcggct cccgacatct tggaccatta
     7381 gctccacagg tatcttcttc cctctagtgg tcataacagc agcttcagct acctctcaat
     7441 ttgcgtatag agcgggtcat cgaagggtta tacactcgta atcgttgcgt tacacacaca
     7501 aaaaaccaac acacatccat cttcgatgga tagcgatttt attatctaac tgctgatcga
     7561 gtgtagccag atctagtaat caattacggg gtcattagtt catagcccat atatggagtt
     7621 ccg
//
