LOCUS       pTwist_CMV_rbIgk        6954 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 6954)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 18, 2025 from SnapGene 8.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6954
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(1175..6954,1..1174)
                     /organism="pTwistCMVrIgk2 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..1495
                     /label=rbIgk2
     misc_feature    1520..2108
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(1991..2002)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2161..2282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     terminator      2397..2443
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    2553..2624
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     promoter        2669..2865
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2716..2851
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2879..3673
                     /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    3684..3908
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4104..4692)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        5438..5542
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5543..5611,5612..6403)
                     /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      6416..6552
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    6697..6768
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     polyA_signal    6826..6874
                     /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 ccgccgcgac cctgtagccc catccgtact
     1201 tctgtttccc ccttctaagg aggaactcac gaccgggacc gccaccatcg tctgcgttgc
     1261 aaacaagttc taccccagtg atatcaccgt cacttggaag gtcgacggaa ctacccaaca
     1321 gagtggcatt gaaaactcca aaaccccgca atctcctgaa gacaacacct attcactgtc
     1381 tagcaccttg tctttgacat ccgcccagta caactcccac tccgtctaca cctgcgaagt
     1441 ggttcagggc agcgccagcc caatcgtcca gtccttcaac cgtggagatt gttgaatcga
     1501 tttctggatc cgcaggccta atcaacctct ggattacaaa atttgtgaaa gattgactgg
     1561 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     1621 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     1681 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     1741 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     1801 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     1861 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     1921 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     1981 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2041 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2101 ctccccgcct gtctagcttg actgactgag atacagcgta ccttcagctc acagacatga
     2161 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     2221 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     2281 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt
     2341 tttaaagcaa gtaaaacctc tacaaatgtg gtattggccc atctctatcg gtatcgtagc
     2401 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgtgcccct cgggccggat
     2461 tgctatctac cgaggcactg gtgtactgat agagccgggc tagatgttaa ctgtcagcta
     2521 tcctgcaggc gatctctcga tttcgatcaa gacattcctt taatggtctt ttctggacac
     2581 cactaggggt cagaagtagt tcatcaaact ttcttccctc cctaatctca ttggttacct
     2641 tgggctatcg aaacttaatt aagcgatctg catctcaatt agtcagcaac catagtcccg
     2701 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2761 cgctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2821 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaggagg tagccaacat
     2881 gattgaacaa gatggattgc acgcaggttc tcccgccgct tgggtggaga ggctattcgg
     2941 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3001 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactcca
     3061 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3121 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3181 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3241 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3301 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3361 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgga tgcccgacgg
     3421 cgaggatctc gtcgtgaccc acggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3481 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3541 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3601 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3661 cgagttcttc tagtatgtaa gccctgtgcc ttctagttgc cagccatctg ttgtttgccc
     3721 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     3781 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     3841 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     3901 ctctatggtt aattaaccag tcaagtcagc tacttggcga gatcgacttg tctgggtttc
     3961 gactacgctc agaattgcgt cagtcaagtt cgatctggtc cttgctattg cacccgttct
     4021 ccgattacga gtttcattta aatcatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4081 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4141 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4201 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4261 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4321 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4381 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4441 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4501 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4561 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4621 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4681 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4741 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4801 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4861 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4921 atagttgcat ttaaatttcc gaactctcca aggccctcgt cggaaaatct tcaaaccttt
     4981 cgtccgatcc atcttgcagg ctacctctcg aacgaactat cgcaagtctc ttggccggcc
     5041 ttgcgccttg gctattgctt ggcagcgcct atcgccaggt attactccaa tcccgaatat
     5101 ccgagatcgg gatcacccga gagaagttca acctacatcc tcaatcccga tctatccgag
     5161 atccgaggaa tatcgaaatc ggggcgcgcc tggtgtaccg agaacgatcc tctcagtgcg
     5221 agtctcgacg atccatatcg ttgcttggca gtcagccagt cggaatccag cttgggaccc
     5281 aggaagtcca atcgtcagat attgtactca agcctggtca cggcagcgta ccgatctgtt
     5341 taaacctaga tattgatagt ctgatcggtc aacgtataat cgagtcctag cttttgcaaa
     5401 catctatcaa gagacaggat cagcaggagg ctttcgccgc ggaaccccta tttgtttatt
     5461 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5521 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5581 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5641 tgctgaagat cagttgggtg cgcgagtggg ttacatcgaa ctggatctca acagcggtaa
     5701 gatccttgag agttttcgcc ccgaagaacg ctttccaatg atgagcactt ttaaagttct
     5761 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5821 acactattct cagaatgact tggttgagta ttcaccagtc acagaaaagc atcttacgga
     5881 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5941 caacttactt ctgacaacga ttggaggacc gaaggagcta accgcttttt tgcacaacat
     6001 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     6061 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca accttgcgta aactattaac
     6121 tggcgaacta cttactctag cttcccggca acagttgata gactggatgg aggcggataa
     6181 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     6241 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     6301 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6361 acagatcgct gagataggtg cctcactgat taagcattgg taaccgattc taggtgcatt
     6421 ggcgcagaaa aaaatgcctg atgcgacgct gcgcgtctta tactcccaca tatgccagat
     6481 tcagcaacgg atacggcttc cccaacttgc ccacttccat acgtgtcctc cttaccagaa
     6541 atttatcctt aagatcgttt aaactcgact ctggctctat cgaatctccg tcgtttcgag
     6601 cttacgcgaa cagccgtggc gctcatttgc tcgtcgggca tcgaatctcg tcagctatcg
     6661 tcagcttacc tttttggcag cgatcgcggc tcccgacatc ttggaccatt agctccacag
     6721 gtatcttctt ccctctagtg gtcataacag cagcttcagc tacctctcaa tttgcgtata
     6781 gagcgggtca tcgaagggtt atacactcgt aatcgttgcg ttacacacac aaaaaaccaa
     6841 cacacatcca tcttcgatgg atagcgattt tattatctaa ctgctgatcg agtgtagcca
     6901 gatctagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccg
//
