LOCUS       pTwist_CMV_rbIgk        6948 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 6948)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 18, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1175..6948,1..1174)
                     /organism="pTwistCMVrIgk1 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..1489
                     /label=rbIgk1
     misc_feature    1514..2102
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(1985..1996)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2155..2276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      2391..2437
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2547..2618
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        2663..2859
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2710..2845
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2873..3667
                     /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    3678..3902
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4098..4686)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5432..5536
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5537..5605,5606..6397)
                     /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      6410..6546
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6691..6762
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    6820..6868
                     /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 ccgcggcgat ccagtggcgc ctactgtgct
     1201 tatctttccc cctgctgccg atcaggtggc caccggcacc gtaaccatcg tctgcgtggc
     1261 caacaagtac tttcctgacg tgacggtaac ctgggaagtc gacggtacga cccagacgac
     1321 cggcatcgag aacagtaaga ctcctcaaaa cagtgcggac tgtacctata atctgtcctc
     1381 taccctgacc ctcacctcaa ctcagtacaa ctctcacaag gagtatacct gcaaagtgac
     1441 ccagggcacg accagtgtcg tacagagttt taaccgcggg gactgctgaa tcgatttctg
     1501 gatccgcagg cctaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     1561 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     1621 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     1681 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     1741 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     1801 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     1861 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt
     1921 tccatggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     1981 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     2041 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     2101 gcctgtctag cttgactgac tgagatacag cgtaccttca gctcacagac atgataagat
     2161 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2221 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2281 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2341 gcaagtaaaa cctctacaaa tgtggtattg gcccatctct atcggtatcg tagcataacc
     2401 ccttggggcc tctaaacggg tcttgagggg ttttttgtgc ccctcgggcc ggattgctat
     2461 ctaccgaggc actggtgtac tgatagagcc gggctagatg ttaactgtca gctatcctgc
     2521 aggcgatctc tcgatttcga tcaagacatt cctttaatgg tcttttctgg acaccactag
     2581 gggtcagaag tagttcatca aactttcttc cctccctaat ctcattggtt accttgggct
     2641 atcgaaactt aattaagcga tctgcatctc aattagtcag caaccatagt cccgccccta
     2701 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatcgctga
     2761 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2821 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag gaggtagcca acatgattga
     2881 acaagatgga ttgcacgcag gttctcccgc cgcttgggtg gagaggctat tcggctatga
     2941 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3001 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tccaggacga
     3061 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3121 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3181 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3241 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3301 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3361 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cggatgcccg acggcgagga
     3421 tctcgtcgtg acccacggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3481 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3541 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     3601 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     3661 cttctagtat gtaagccctg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     3721 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3781 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3841 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3901 ggttaattaa ccagtcaagt cagctacttg gcgagatcga cttgtctggg tttcgactac
     3961 gctcagaatt gcgtcagtca agttcgatct ggtccttgct attgcacccg ttctccgatt
     4021 acgagtttca tttaaatcat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4081 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4141 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4201 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4261 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4321 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4381 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4441 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4501 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4561 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4621 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4681 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4741 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4801 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4861 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4921 gcatttaaat ttccgaactc tccaaggccc tcgtcggaaa atcttcaaac ctttcgtccg
     4981 atccatcttg caggctacct ctcgaacgaa ctatcgcaag tctcttggcc ggccttgcgc
     5041 cttggctatt gcttggcagc gcctatcgcc aggtattact ccaatcccga atatccgaga
     5101 tcgggatcac ccgagagaag ttcaacctac atcctcaatc ccgatctatc cgagatccga
     5161 ggaatatcga aatcggggcg cgcctggtgt accgagaacg atcctctcag tgcgagtctc
     5221 gacgatccat atcgttgctt ggcagtcagc cagtcggaat ccagcttggg acccaggaag
     5281 tccaatcgtc agatattgta ctcaagcctg gtcacggcag cgtaccgatc tgtttaaacc
     5341 tagatattga tagtctgatc ggtcaacgta taatcgagtc ctagcttttg caaacatcta
     5401 tcaagagaca ggatcagcag gaggctttcg ccgcggaacc cctatttgtt tatttttcta
     5461 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     5521 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     5581 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     5641 agatcagttg ggtgcgcgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5701 tgagagtttt cgccccgaag aacgctttcc aatgatgagc acttttaaag ttctgctatg
     5761 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5821 ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta cggatggcat
     5881 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5941 acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     6001 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     6061 gcgtgacacc acgatgcctg tagcaatggc aacaaccttg cgtaaactat taactggcga
     6121 actacttact ctagcttccc ggcaacagtt gatagactgg atggaggcgg ataaagttgc
     6181 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     6241 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     6301 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     6361 cgctgagata ggtgcctcac tgattaagca ttggtaaccg attctaggtg cattggcgca
     6421 gaaaaaaatg cctgatgcga cgctgcgcgt cttatactcc cacatatgcc agattcagca
     6481 acggatacgg cttccccaac ttgcccactt ccatacgtgt cctccttacc agaaatttat
     6541 ccttaagatc gtttaaactc gactctggct ctatcgaatc tccgtcgttt cgagcttacg
     6601 cgaacagccg tggcgctcat ttgctcgtcg ggcatcgaat ctcgtcagct atcgtcagct
     6661 tacctttttg gcagcgatcg cggctcccga catcttggac cattagctcc acaggtatct
     6721 tcttccctct agtggtcata acagcagctt cagctacctc tcaatttgcg tatagagcgg
     6781 gtcatcgaag ggttatacac tcgtaatcgt tgcgttacac acacaaaaaa ccaacacaca
     6841 tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta gccagatcta
     6901 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
