LOCUS       pTwist_CMV_bispe        7314 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 7314)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 4, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7314
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1175..7314,1..1174)
                     /organism="bispecifichIgG1(stumphole) 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..1855
                     /label=bispecific hIgG1 stump hole
     misc_feature    1880..2468
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2351..2362)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2521..2642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      2757..2803
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2913..2984
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        3029..3225
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3076..3211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3239..4033
                     /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    4044..4268
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4464..5052)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5798..5902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5903..5971,5972..6763)
                     /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      6776..6912
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    7057..7128
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    7186..7234
                     /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 ccgcgataag actcacacct gtccgccatg
     1201 ccccgccccg gaacttctgg gtggccctag tgtgtttctg ttccctccca aacctaagga
     1261 cacactcatg attagcagaa ctccagaagt cacgtgcgtg gtcgtggatg tgtcccacga
     1321 ggacccagag gtgaagttta actggtacgt agatggcgtg gaggttcaca acgcaaagac
     1381 taagccgcgc gaagagcagt acaactccac atacagagtc gtgtctgtgc tcaccgtgct
     1441 gcatcaggac tggctgaatg gcaaagagta caagtgcaaa gtgtccaaca aggctctgcc
     1501 cgctccaatc gagaagacga tctccaaggc caagggtcag ccccgtgagc cccaggtgta
     1561 taccctgccg ccctctcgtg aggaaatgac caagaaccag gtgagcctga gttgtgcagt
     1621 gaagggcttc tatccttccg atatcgccgt cgagtgggaa agcaacgggc agccagagaa
     1681 taactacaag accacacccc ctgtgctcga ctccgacggg tcatttttcc tggtatctaa
     1741 actcaccgtt gacaagagca ggtggcagca aggaaacgtg ttctcctgct ccgttatgca
     1801 cgaggccctg cataaccact acacgcagaa atccctcagc ctgagccccg ggtgaatcga
     1861 tttctggatc cgcaggccta atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1921 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1981 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2041 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2101 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2161 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2221 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2281 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2341 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2401 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2461 ctccccgcct gtctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     2521 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2581 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2641 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2701 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     2761 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     2821 tgctatctac cgaggcactg gtgtactgat agagccgggc tagatgttaa ctgtcagcta
     2881 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     2941 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     3001 tgggctatcg aaacttaatt aagcgatctg catctcaatt agtcagcaac catagtcccg
     3061 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3121 cgctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3181 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaggagg tagccaacat
     3241 gattgaacaa gatggattgc acgcaggttc tcccgccgct tgggtggaga ggctattcgg
     3301 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3361 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactcca
     3421 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3481 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3541 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3601 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3661 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3721 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgga tgcccgacgg
     3781 cgaggatctc gtcgtgaccc acggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3841 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3901 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3961 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4021 cgagttcttc tagtatgtaa gccctgtgcc ttctagttgc cagccatctg ttgtttgccc
     4081 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4141 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4201 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4261 ctctatggtt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
     4321 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
     4381 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4441 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4501 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4561 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4621 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4681 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4741 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4801 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4861 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4921 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4981 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5041 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5101 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5161 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5221 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5281 atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt
     5341 cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc
     5401 ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat
     5461 ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag
     5521 atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg
     5581 agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc
     5641 aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt
     5701 taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa
     5761 catctatcaa gagacaggat cagcaggagg ctttcgccgc ggaaccccta tttgtttatt
     5821 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5881 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5941 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     6001 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     6061 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     6121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     6181 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     6241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     6301 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     6361 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     6481 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     6541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6721 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     6781 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     6841 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     6901 atttatcctt aagatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     6961 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     7021 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     7081 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa tttgcgtata
     7141 gagcgggtca tcgaagggtt atacactcgt aatcgttgcg ttacacacac aaaaaaccaa
     7201 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     7261 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
