LOCUS       sequence-231769-        5653 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 5653)
  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 5653)
  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-KPNA2
FEATURES             Location/Qualifiers
     source          1..5653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        143..522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..726
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     677..697
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     723..747
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          862..994
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     977..996
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1039..1057
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        1063..1081
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1063..1080
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      1112..1121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2772..2893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2783..2802)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2837..2856
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3074..3529
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3161..3180)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3371..3392
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3596..3615)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3715..3737
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3775..3793)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3861..3965
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3966..4034,4035..4826)
                     /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(4184..4203)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4997..5585
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5486..5505
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gatcttcaat attggccatt agccatatta ttcattggtt atatagcata aatcaatatt
       61 ggctattggc cattgcatac gttgtatcta tatcataata tgtacattta tattggctca
      121 tgtccaatat gaccgccatg ttggcattga ttattgacta gttattaata gtaatcaatt
      181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      541 tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat
      661 aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      721 agagctcgtt tagtgaaccg tcagatcact agaagcttta ttgcggtagt ttatcacagt
      781 taaattgcta acgcagtcag tgcttctgac acaacagtct cgaacttaag ctgcagaagt
      841 tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta aggagaccaa
      901 tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc acctattggt
      961 cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt caattacagc
     1021 tcttaaggct agagtactta atacgactca ctataggcta gcatttaggt gacactatag
     1081 aatacaagct acttgttctt tttgcactcg agccaccatg gccagcatga ccggcggcca
     1141 gcagatgggc gaattctcca ccaacgagaa tgctaataca ccagctgccc gtcttcacag
     1201 attcaagaac aagggaaaag acagtacaga aatgaggcgt cgcagaatag aggtcaatgt
     1261 ggagctgagg aaagctaaga aggatgacca gatgctgaag aggagaaatg taagctcatt
     1321 tcctgatgat gctacttctc cgctgcagga aaaccgcaac aaccagggca ctgtaaattg
     1381 gtctgttgat gacattgtca aaggcataaa tagcagcaat gtggaaaatc agctccaagc
     1441 tactcaagct gccaggaaac tactttccag agaaaaacag ccccccatag acaacataat
     1501 ccgggctggt ttgattccga aatttgtgtc cttcttgggc agaactgatt gtagtcccat
     1561 tcagtttgaa tctgcttggg cactcactaa cattgcttct gggacatcag aacaaaccaa
     1621 ggctgtggta gatggaggtg ccatcccagc attcatttct ctgttggcat ctccccatgc
     1681 tcacatcagt gaacaagctg tctgggctct aggaaacatt gcaggtgatg gctcagtgtt
     1741 ccgagacttg gttattaagt acggtgcagt tgacccactg ttggctctcc ttgcagttcc
     1801 tgatatgtca tctttagcat gtggctactt acgtaatctt acctggacac tttctaatct
     1861 ttgccgcaac aagaatcctg cacccccgat agatgctgtt gagcagattc ttcctacctt
     1921 agttcggctc ctgcatcatg atgatccaga agtattagca gatacctgct gggctatttc
     1981 ctaccttact gatggtccaa atgaacgaat tggcatggtg gtgaaaacag gagttgtgcc
     2041 ccaacttgtg aagcttctag gagcttctga attgccaatt gtgactcctg ccctaagagc
     2101 catagggaat attgtcactg gtacagatga acagactcag gttgtgattg atgcaggagc
     2161 actcgccgtc tttcccagcc tgctcaccaa ccccaaaact aacattcaga aggaagctac
     2221 gtggacaatg tcaaacatca cagccggccg ccaggaccag atacagcaag ttgtgaatca
     2281 tggattagtc ccattccttg tcagtgttct ctctaaggca gattttaaga cacaaaagga
     2341 agctgtgtgg gccgtgacca actataccag tggtggaaca gttgaacaga ttgtgtacct
     2401 tgttcactgt ggcataatag aaccgttgat gaacctctta actgcaaaag ataccaagat
     2461 tattctggtt atcctggatg ccatttcaaa tatctttcag gctgctgaga aactaggtga
     2521 aactgagaaa cttagtataa tgattgaaga atgtggaggc ttagacaaaa ttgaagctct
     2581 acaaaaccat gaaaatgagt ctgtgtataa ggcttcgtta agcttaattg agaagtattt
     2641 ctctgtagag gaagaggaag atcaaaacgt tgtaccagaa actacctctg aaggctacac
     2701 tttccaagtt caggatgggg ctcctgggac ctttaacttt tagcccgggc ggccgcttcg
     2761 agcagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     2821 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     2881 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat
     2941 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc
     3001 gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     3061 gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3121 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3181 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3241 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3301 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3361 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3421 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3481 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc ctgatgcggt
     3541 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     3601 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3661 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3721 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3781 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3841 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3901 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3961 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4021 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4081 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4141 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4201 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4261 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4321 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4381 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4441 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4501 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4561 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4621 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4681 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4741 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4801 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4861 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4921 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4981 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5041 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5101 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5161 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5221 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5281 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5341 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5401 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5461 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5521 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5581 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5641 acatggctcg aca
//
