LOCUS       sequence-309028-        6470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses API5 in E.coli cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6470)
  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 6470)
  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: pGEX-API5
FEATURES             Location/Qualifiers
     source          1..6470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(196..201,202..217,218..224)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    232..248
                     /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)."
     CDS             271..924
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     882..904
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             931..948
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2533..2555)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2700..2718)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2786..2890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2891..2959,2960..3751)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3109..3128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3922..4510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4411..4430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4664..4681
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4754..4831
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4832..5914
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4851..4870)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5963..5968,5969..5986,5987..5993)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6001..6017
                     /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)."
     primer_bind     6025..6041
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6025..6041
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6037..6470,1..94)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6057..6074)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6057..6073)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6066..6088)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg tcaggcagcc
       61 atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa
      121 ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt ctggcaaata
      181 ttctgaaatg agctgttgac aattaatcat cggctcgtat aatgtgtgga attgtgagcg
      241 gataacaatt tcacacagga aacagtattc atgtccccta tactaggtta ttggaaaatt
      301 aagggccttg tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag
      361 catttgtatg agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg
      421 gagtttccca atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc
      481 atcatacgtt atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca
      541 gagatttcaa tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca
      601 tatagtaaag actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg
      661 aaaatgttcg aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat
      721 cctgacttca tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg
      781 gatgcgttcc caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat
      841 aagtacttga aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt
      901 ggtggtggcg accatcctcc aaaatcggat ctggttccgc gtggatcccc gaattcaatg
      961 ccgacagtag aggagcttta ccgcaattat ggcatcctgg ccgatgccac ggagcaagtg
     1021 ggccagcata aagatgccta tcaagtgata ttggatggtg tgaaaggtgg tactaaggaa
     1081 aagcgattag cagctcaatt tattccgaaa ttctttaagc attttccaga attggctgat
     1141 tctgctatca atgcacagtt agacctctgt gaggatgaag atgtatctat tcgacgtcaa
     1201 gcaattaaag aactgcctca atttgccact ggagaaaatc ttcctcgagt ggcagatata
     1261 ctaacgcaac ttttgcagac agatgactct gcagaattta acctagtgaa caatgcccta
     1321 ttaagtatat ttaaaatgga tgcaaaaggg actttaggtg ggttgttcag ccaaatactt
     1381 caaggagagg acattgttag agaacgagca attaaattcc tttctacaaa acttaagact
     1441 ttaccagatg aagtcttaac aaaggaagtg gaagagctta tactaactga atccaaaaag
     1501 gtcctagaag atgtgactgg tgaagaattt gttctattta tgaagatact gtctgggtta
     1561 aaaagcttac agacagtgag tggaagacag caacttgtag agttggtggc tgaacaggcc
     1621 gacctagaac agaccttcaa tccctcggat cctgactgtg tggacaggct cttacagtgc
     1681 actcggcagg cagtacccct cttctctaaa aatgtccatt ccacaaggtt tgtgacatat
     1741 ttctgtgagc aggttctccc taacctcggt accttgacta ccccagtgga aggtcttgat
     1801 atacagttgg aggtattgaa attgttggcg gagatgagtt cattttgtgg tgacatggaa
     1861 aaactagaaa caaatttaag gaaactattt gataagttat tggaatacat gcccctccct
     1921 ccagaagagg cagaaaatgg agagaatgct ggtaatgaag aacccaagct acagttcagt
     1981 tatgtggaat gtttgttgta cagttttcac cagttgggcc gaaaacttcc agatttctta
     2041 acagccaaac tgaatgcaga aaagctcaaa gatttcaaaa tcaggctgca gtactttgca
     2101 cggggcctgc aagtttatat cagacaactt cgcttagctc tccagggtaa aacgggtgag
     2161 gccttaaaaa cagaagagaa caagattaaa gtcgttgcat tgaaaataac aaacaatatc
     2221 aatgttttaa tcaaggatct cttccacatt cctccttctt ataagagcac agtaacacta
     2281 tcctggaaac ctgtacaaaa ggttgagatt gggcaaaaga gagccagtga agatacaact
     2341 tcaggttcac cacccaagaa atcttcagca ggaccaaaaa gagatgccag gcagatttat
     2401 aaccctccca gtgggaaata tagcagcaat ttgggcaact ttaattatga gaggagcctt
     2461 caggggaagt aagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
     2521 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2581 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     2641 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
     2701 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2761 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2821 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2881 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2941 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3001 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3061 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3121 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3181 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3241 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3301 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3361 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3421 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3481 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3541 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3601 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3661 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3721 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3781 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3841 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3901 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3961 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4021 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4081 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4141 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4201 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4261 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4321 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4381 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4441 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4501 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4561 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4621 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4681 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     4741 ccgcataaat tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     4801 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     4861 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     4921 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     4981 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     5041 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     5101 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     5161 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     5221 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     5281 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     5341 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     5401 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     5461 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     5521 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     5581 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     5641 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     5701 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     5761 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     5821 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     5881 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     5941 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     6001 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg
     6061 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     6121 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     6181 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
     6241 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
     6301 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
     6361 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     6421 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga
//
