LOCUS       sequence-309044-        7798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses API5 in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  AUTHORS   Bong SM, Bae SH, Song B, Gwak H, Yang SW, Kim S, Nam S, Rajalingam 
            K, Oh SJ, Kim TW, Park S, Jang H, Lee BI
  TITLE     Regulation of mRNA export through API5 and nuclear FGF2 interaction.
  JOURNAL   Nucleic Acids Res. 2020 Jun 19;48(11):6340-6352. doi: 
            10.1093/nar/gkaa335.
  PUBMED    32383752
REFERENCE   2  (bases 1 to 7798)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2020-06-19- 19"; volume: "48"; issue: 
            "11"; pages: "6340-6352"
COMMENT     Sequence Label: pCAG-F-API5
FEATURES             Location/Qualifiers
     source          1..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(8..799,800..868))
                     /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     631..650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1461..1595
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        1609..1988
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1990..2265
                     /label=chicken beta-actin promoter
     intron          2266..3282
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3206..3228
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3290..3309
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3337..3346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3346..3369
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    4920..5493
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4943..5495
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5087..5104)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5314..5333
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5495..5893
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(5913..5946)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(6023..6040)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6029..6253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6300..6316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6300..6316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6313..6335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6324..6340
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6348..6354,6355..6372,6373..6378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6393..6414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6473..6668
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6519..6654
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6674..6808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6876..6893)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7047..7635)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7127..7146)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
       61 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
      121 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
      181 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
      241 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
      301 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
      361 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
      421 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
      481 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
      541 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
      601 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
      661 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
      721 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
      781 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
      841 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatgaccg ctttggaaaa
      901 acaaagactg tatttcctgg aaattaatgt ttattcaata aactgtgtat tcagctatat
      961 tcacatagtg gtgaggctga aatgaggcgg gaagaggcgg ttggggctta attatatcaa
     1021 tttgggtggc cccacagcgc ctccaaggcg ccagtcctgt tttgacaagt tgcctctgga
     1081 agcctctctt ctttttctcc agagtaagcg gaggccaggg gcccccggcc tctgcttaat
     1141 actaaaaaaa acagctgttg tcatagtaat gattgggtgg aaacattcca ggcctgggtg
     1201 gagaggcttt ttgcttcctc ttgcaaaacc acactgacat tccaggcctg ggtggagagg
     1261 ctttttgctt cctcttgcaa aaccacactg ccctctggag ggcagttgcc tagcaactaa
     1321 ttaaaagagg atgtcgcacg gccagctgcg gtcagttagt cacttcctgc ttaactgact
     1381 tgacattttc tattttaaga gtcgggagga aaattactgt gttggaggcc ctccgccatc
     1441 ttctgaagct gaatcgaatt aacttgttta ttgcagctta taatggttac aaataaagca
     1501 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     1561 ccaaactcat caatgtatct tatcatgtct ggatctgata tcatcgtcga cattgattat
     1621 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     1681 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     1741 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     1801 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     1861 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1921 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     1981 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     2041 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     2101 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg
     2161 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg
     2221 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcgggagt cgctgcgcgc
     2281 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
     2341 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
     2401 cgcttggttt aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc
     2461 cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg
     2521 gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg
     2581 gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg
     2641 tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga
     2701 gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt
     2761 gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc
     2821 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc
     2881 ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg
     2941 gcgagccgca gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg
     3001 tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc
     3061 ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg
     3121 ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc
     3181 cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga
     3241 gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
     3301 gttattgtgc tgtctcatca ttttggcaaa gaattagcca ccatggatta caaggatgac
     3361 gacgataagc tcgagcccgg gatgccgaca gtagaggagc tttaccgcaa ttatggcatc
     3421 ctggccgatg ccacggagca agtgggccag cataaagatg cctatcaagt gatattggat
     3481 ggtgtgaaag gtggtactaa ggaaaagcga ttagcagctc aatttattcc gaaattcttt
     3541 aagcattttc cagaattggc tgattctgct atcaatgcac agttagacct ctgtgaggat
     3601 gaagatgtat ctattcgacg tcaagcaatt aaagaactgc ctcaatttgc cactggagaa
     3661 aatcttcctc gagtggcaga tatactaacg caacttttgc agacagatga ctctgcagaa
     3721 tttaacctag tgaacaatgc cctattaagt atatttaaaa tggatgcaaa agggacttta
     3781 ggtgggttgt tcagccaaat acttcaagga gaggacattg ttagagaacg agcaattaaa
     3841 ttcctttcta caaaacttaa gactttacca gatgaagtct taacaaagga agtggaagag
     3901 cttatactaa ctgaatccaa aaaggtccta gaagatgtga ctggtgaaga atttgttcta
     3961 tttatgaaga tactgtctgg gttaaaaagc ttacagacag tgagtggaag acagcaactt
     4021 gtagagttgg tggctgaaca ggccgaccta gaacagacct tcaatccctc ggatcctgac
     4081 tgtgtggaca ggctcttaca gtgcactcgg caggcagtac ccctcttctc taaaaatgtc
     4141 cattccacaa ggtttgtgac atatttctgt gagcaggttc tccctaacct cggtaccttg
     4201 actaccccag tggaaggtct tgatatacag ttggaggtat tgaaattgtt ggcggagatg
     4261 agttcatttt gtggtgacat ggaaaaacta gaaacaaatt taaggaaact atttgataag
     4321 ttattggaat acatgcccct ccctccagaa gaggcagaaa atggagagaa tgctggtaat
     4381 gaagaaccca agctacagtt cagttatgtg gaatgtttgt tgtacagttt tcaccagttg
     4441 ggccgaaaac ttccagattt cttaacagcc aaactgaatg cagaaaagct caaagatttc
     4501 aaaatcaggc tgcagtactt tgcacggggc ctgcaagttt atatcagaca acttcgctta
     4561 gctctccagg gtaaaacggg tgaggcctta aaaacagaag agaacaagat taaagtcgtt
     4621 gcattgaaaa taacaaacaa tatcaatgtt ttaatcaagg atctcttcca cattcctcct
     4681 tcttataaga gcacagtaac actatcctgg aaacctgtac aaaaggttga gattgggcaa
     4741 aagagagcca gtgaagatac aacttcaggt tcaccaccca agaaatcttc agcaggacca
     4801 aaaagagatg ccaggcagat ttataaccct cccagtggga aatatagcag caatttgggc
     4861 aactttaatt atgagaggag ccttcagggg aagtaagcgg ccgcgaattc cgcccccccc
     4921 ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     4981 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     5041 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     5101 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     5161 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     5221 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     5281 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     5341 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     5401 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     5461 gacgtggttt tcctttgaaa aacacgatga taatatggcc aagcctttgt ctcaagaaga
     5521 atccaccctc attgaaagag caacggctac aatcaacagc atccccatct ctgaagacta
     5581 cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg tcaatgtata
     5641 tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg ctgctgcggc
     5701 agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca tcttgagccc
     5761 ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag ccatagtgaa
     5821 ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct ctggttatgt
     5881 gtgggagggc taagaattcg tcgagggacc taataacttc gtatagcata cattatacga
     5941 agttatacat gtttaagggt tccggttcca ctaggtacaa ttcgatatca agcttatcga
     6001 tgatctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     6061 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     6121 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     6181 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     6241 gtgggctcta tggcttctga ggcggaaaga acctgcagcc caagcttggc gtaatcatgg
     6301 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6361 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6421 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     6481 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6541 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6601 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6661 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6721 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6781 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     6841 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6901 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6961 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7021 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7081 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7141 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7201 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7261 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7321 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7381 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7441 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7501 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7561 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7621 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7681 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7741 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggt
//
