LOCUS       sequence-309047-        6903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses FGF2 in mammalian cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6903)
  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 6903)
  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-HA-LMW-FGF2
FEATURES             Location/Qualifiers
     source          1..6903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(12..803,804..872))
                     /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     635..654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    1465..1599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     enhancer        1613..1992
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1994..2271
                     /label=chicken beta-actin promoter
     intron          2272..3288
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3212..3234
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3296..3315
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3352..3378
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    join(3865..4439,4440..4442,4443..4451)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4032..4049)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4259..4278
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4452..5051
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4452..4471)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4948..4968
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(5132..5149)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5138..5362
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5409..5425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5409..5425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5422..5444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5433..5449
                     /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(5457..5463,5464..5481,5482..5487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5502..5523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5582..5777
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5628..5763
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5783..5917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5985..6002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6156..6744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6236..6255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
       61 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
      121 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
      181 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
      241 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
      301 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
      361 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
      421 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
      481 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
      541 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
      601 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
      661 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
      721 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
      781 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
      841 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaatg accgctttgg
      901 aaaaacaaag actgtatttc ctggaaatta atgtttattc aataaactgt gtattcagct
      961 atattcacat agtggtgagg ctgaaatgag gcgggaagag gcggttgggg cttaattata
     1021 tcaatttggg tggccccaca gcgcctccaa ggcgccagtc ctgttttgac aagttgcctc
     1081 tggaagcctc tcttcttttt ctccagagta agcggaggcc aggggccccc ggcctctgct
     1141 taatactaaa aaaaacagct gttgtcatag taatgattgg gtggaaacat tccaggcctg
     1201 ggtggagagg ctttttgctt cctcttgcaa aaccacactg acattccagg cctgggtgga
     1261 gaggcttttt gcttcctctt gcaaaaccac actgccctct ggagggcagt tgcctagcaa
     1321 ctaattaaaa gaggatgtcg cacggccagc tgcggtcagt tagtcacttc ctgcttaact
     1381 gacttgacat tttctatttt aagagtcggg aggaaaatta ctgtgttgga ggccctccgc
     1441 catcttctga agctgaatcg aattaacttg tttattgcag cttataatgg ttacaaataa
     1501 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     1561 ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatatcatcg tcgacattga
     1621 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     1681 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     1741 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     1801 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     1861 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     1921 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     1981 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     2041 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     2101 gggggggggg gggggcgcgc gccrggcggg gcggggcggg gcgaggggcg gggcggggcg
     2161 aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg
     2221 gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct
     2281 gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
     2341 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
     2401 aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag
     2461 gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg
     2521 tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc
     2581 ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc
     2641 ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg
     2701 gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc
     2761 cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg
     2821 ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct
     2881 cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga
     2941 ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt
     3001 cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc
     3061 gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt
     3121 gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac
     3181 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
     3241 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
     3301 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tagccaccat gtacccatac
     3361 gatgttccag attacgctct cgaggcggct ggctctatca ccactctgcc ggccctgccg
     3421 gaagatggcg gcagcggcgc gttcccgccg ggccatttta aggatccgaa gcgcctgtac
     3481 tgcaaaaacg gcggcttttt cctgcgcatt cacccggatg gccgtgtgga cggtgtgcgt
     3541 gagaaaagcg acccgcacat caaactgcaa ctgcaagcgg aggaacgtgg tgtagtgagc
     3601 atcaagggtg tatgcgctaa ccgttatctg gcgatgaaag aagacggccg cctgctggcg
     3661 agcaaatgcg tcaccgacga atgtttcttc ttcgaacgcc tggagtccaa caactacaac
     3721 acgtatcgct cccgcaaata cacctcttgg tacgtggctc tgaaacgcac cggccagtac
     3781 aaactgggct ccaaaaccgg ccctggccag aaagcgatcc tgttcctgcc gatgtccgcg
     3841 aaatcctaag aattccgccc cccccccctc tccctccccc ccccctaacg ttactggccg
     3901 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     3961 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     4021 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     4081 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     4141 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     4201 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     4261 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     4321 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     4381 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataata
     4441 tggccacaac catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
     4501 ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
     4561 tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
     4621 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
     4681 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
     4741 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
     4801 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
     4861 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
     4921 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
     4981 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
     5041 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
     5101 atgcatcgat gatctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca
     5161 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     5221 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     5281 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     5341 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa cctgcagccc aagcttggcg
     5401 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5461 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5521 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     5581 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     5641 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     5701 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     5761 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     5821 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     5881 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     5941 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     6001 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     6061 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6121 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6181 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6241 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6301 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6361 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6421 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6481 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6541 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6601 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6661 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6721 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6781 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6841 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6901 act
//
