LOCUS       sequence-232280-        5641 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 5641)
  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 5641)
  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-KPNA4
FEATURES             Location/Qualifiers
     source          1..5641
                     /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    2760..2881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2771..2790)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2825..2844
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3062..3517
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3149..3168)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3359..3380
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3584..3603)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3703..3725
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3763..3781)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3849..3953
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3954..4022,4023..4814)
                     /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(4172..4191)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4985..5573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5474..5493
                     /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 gaattcacgc gtggtaccgc ggacaacgag aaactggaca accaacggct
     1201 caagaatttc aagaacaaag gccgcgactt ggagactatg agaagacaac gaaatgaagt
     1261 tgtagttgaa ttaaggaaga ataaaagaga tgaacatctc ttaaagagaa ggaatgtacc
     1321 acatgaagat atctgtgaag actctgatat agatggtgat tatagagtgc aaaatacctc
     1381 tctagaagct attgttcaaa atgcttcaag tgataaccaa ggaattcaat taagtgcagt
     1441 tcaagctgct aggaagcttt tgtccagtga tcgaaatcca ccaattgatg acttaataaa
     1501 atctggaata ttgcccattt tagtccattg tcttgaaaga gatgacaatc cttctttaca
     1561 gtttgaagct gcatgggctt tgacaaacat tgcatctgga acttctgaac aaactcaagc
     1621 agtagttcag tccaatgctg tgccactttt cctgaggctt ctccattcac cccatcagaa
     1681 tgtctgtgag caagcagtgt gggcattggg aaatatcata ggtgatgggc cccagtgtag
     1741 agattatgtc ataagtcttg gagttgtgaa acctttactt tccttcataa gtccatctat
     1801 tcctataaca ttcttaagaa atgttacttg ggttatggtc aacttatgtc gccacaaaga
     1861 cccaccacca ccaatggaaa ccattcagga gattcttcca gccctttgtg ttttaattca
     1921 tcacacagat gtaaatatac tggtagacac agtctgggcc ctctcttacc ttactgatgc
     1981 tggcaatgaa caaatacaga tggtaataga ctctggaata gttcctcatt tggttcctct
     2041 gctcagccac caggaagtta aagttcagac tgctgcactt agagctgtgg gcaacattgt
     2101 tactggaact gatgagcaaa cacaagtagt tttgaactgt gatgctcttt cacacttccc
     2161 agcactcctg acacatccca aagagaaaat taataaagaa gcagtgtggt tcctctccaa
     2221 catcactgca ggaaatcagc agcaggtaca ggcagtaatt gatgccaatc ttgtaccaat
     2281 gataatacac cttttggata agggggattt tggcactcaa aaagaagctg cttgggccat
     2341 aagtaactta acaattagtg gaaggaaaga tcaagtggct taccttatcc aacaaaatgt
     2401 tatcccacct ttttgcaact tgctgactgt aaaagatgca caagttgtgc aagtagtact
     2461 cgatggacta agtaatatat taaaaatggc tgaagatgag gcagaaacca taggcaatct
     2521 tatagaagaa tgtggagggc tggagaaaat tgaacaactt caaaatcatg aaaatgaaga
     2581 catctacaaa ttggcctatg agatcattga tcagttcttc tcttcagatg atattgatga
     2641 agaccctagc cttgttccag aggcaattca aggcggaaca tttggtttca attcatctgc
     2701 caatgtacca acagaagggt tccagtttta gcccgggcgg ccgcttcgag cagacatgat
     2761 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     2821 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     2881 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     2941 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatccgg gctggcgtaa
     3001 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     3061 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3121 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3181 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3241 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3301 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3361 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3421 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3481 aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta
     3541 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     3601 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3661 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3721 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     3781 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     3841 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3901 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3961 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     4021 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     4081 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     4141 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     4201 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     4261 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     4321 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     4381 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     4441 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     4501 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     4561 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     4621 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     4681 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4741 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4801 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4861 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4921 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4981 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5041 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5101 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     5161 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5221 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5281 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5341 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5401 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5461 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5521 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5581 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac
     5641 a
//
