LOCUS       sequence-232281-        5674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5674)
  AUTHORS   Kelley JB, Talley AM, Spencer A, Gioeli D, Paschal BM
  TITLE     Karyopherin alpha7 (KPNA7), a divergent member of the importin alpha
            family of nuclear import receptors.
  JOURNAL   BMC Cell Biol. 2010 Aug 11;11:63. doi: 10.1186/1471-2121-11-63.
  PUBMED    20701745
REFERENCE   2  (bases 1 to 5674)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "BMC Cell 
            Biol."; date: "2010-08-11"; volume: "11:63. doi"; pages: " 
            10.1186/1471-2121-11-63"
COMMENT     Sequence Label: pCMVTNT-T7-KPNA6
FEATURES             Location/Qualifiers
     source          1..5674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        62..365
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        366..569
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     520..540
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     566..590
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          705..837
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     820..839
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        882..900
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        906..924
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     906..923
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      955..964
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2636..2757
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2647..2666)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2701..2720
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2938..3393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3025..3044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3235..3256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3460..3479)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3579..3601
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3639..3657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3725..3829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3830..3898,3899..4690)
                     /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"
     primer_bind     complement(4048..4067)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4861..5449
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5350..5369
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
       61 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      121 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      181 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      241 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      301 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      361 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
      421 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      481 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg
      541 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc actagaagct
      601 ttattgcggt agtttatcac agttaaattg ctaacgcagt cagtgcttct gacacaacag
      661 tctcgaactt aagctgcaga agttggtcgt gaggcactgg gcaggtaagt atcaaggtta
      721 caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga agactcttgc
      781 gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc tccacaggtg
      841 tccactccca gttcaattac agctcttaag gctagagtac ttaatacgac tcactatagg
      901 ctagcattta ggtgacacta tagaatacaa gctacttgtt ctttttgcac tcgagccacc
      961 atggccagca tgaccggcgg ccagcagatg ggcgaattcg agaccatggc gagcccaggg
     1021 aaagacaatt atcgaatgaa gagctataag aacaatgctc taaaccctga agaaatgaga
     1081 cgaagaagag aggaagaggg cattcagctc cggaagcaga agcgagagca acaacttttt
     1141 aaacggagaa atgtggagct gattaatgaa gaagctgcca tgttcgatag tcttctcatg
     1201 gactcttatg tgagctctac cactggggag agtgtgatca caagagagat ggtggagatg
     1261 ctcttttctg atgattctga cctgcagtta gcaaccacac agaaattccg gaaactgctc
     1321 tccaaagagc ctagtcctcc aatagatgaa gttatcaaca ctccaagagt ggtggatcgg
     1381 ttcgtggagt ttctgaagag gaatgagaat tgtacattac agtttgaagc tgcctgggct
     1441 ctaacgaata ttgcctctgg aacctctcag cagaccaaaa ttgtcattga agcaggggct
     1501 gtccccattt ttatagagct gcttaattca gactttgagg atgttcagga acaggcagtc
     1561 tgggcactgg gaaacatagc tggagatagc tctgtttgcc gagattacgt cttgaactgt
     1621 tccatcctta atcctttgtt aacactcctt accaagtcca cacgactgac gatgacacgg
     1681 aatgcagtct gggccctgtc aaatctctgc cgagggaaaa acccaccccc agagtttgca
     1741 aaggtctctc cttgtttgcc tgtactgtct cgcctactct tcagcagcga ctcggacttg
     1801 ctggcagatg cttgctgggc cctttcttat ctgtctgatg gccccaatga gaagatccag
     1861 gcagtcatag actccggagt ctgccggaga ttggtagagc tgctgatgca caatgattac
     1921 aaagtggctt ctcctgccct gagagccgtg ggtaacatcg tcactgggga tgacatccag
     1981 acccaggtca ttcttaactg ttcagcccta ccttgccttc tccacttgtt gagcagtccc
     2041 aaggagtcaa tccggaagga agcttgctgg actatttcaa atattactgc tggcaacagg
     2101 gctcaaatac aggctgttat agatgcaaat atcttccctg tgttgatcga aatccttcag
     2161 aaagcagagt ttcgtacaag gaaagaggca gcctgggcca tcaccaatgc cacatcagga
     2221 ggaacccctg agcagatcag gtacctggtc tcactgggct gcatcaaacc cctatgtgac
     2281 ttgctgactg taatggattc gaagattgtg caagtggccc tcaatggact ggagaacatc
     2341 ctgcggcttg gagagcaaga gggcaagcgc agtggctcag gggtcaatcc ttattgtggc
     2401 ctcatagagg aagcctatgg cttggataaa attgagtttc tccagagcca cgagaaccag
     2461 gagatctacc agaaggcctt cgacctcatt gagcactact ttggtgtaga agacgatgat
     2521 agcagcctgg ctccccaagt cgatgaaacg caacagcagt tcatcttcca gcagcctgag
     2581 gcccccatgg agggcttcca gctataaccc gggcggccgc ttcgagcaga catgataaga
     2641 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2701 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2761 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     2821 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atccgggctg gcgtaatagc
     2881 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     2941 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3001 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3061 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3121 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3181 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3241 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3301 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3361 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     3421 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     3481 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3541 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3601 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3661 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3781 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3841 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3901 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3961 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4021 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4081 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4141 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4201 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4261 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4321 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4381 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4441 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4501 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4561 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4621 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4681 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4741 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4801 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4861 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4921 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4981 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     5041 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5101 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5161 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5221 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5281 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5341 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5401 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5461 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
     5521 cttcaatatt ggccattagc catattattc attggttata tagcataaat caatattggc
     5581 tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt
     5641 ccaatatgac cgccatgttg gcattgatta ttga
//
