LOCUS       sequence-350798-        5563 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 5563)
  AUTHORS   Zhou BP, Deng J, Xia W, Xu J, Li YM, Gunduz M, Hung MC
  TITLE     Dual regulation of Snail by GSK-3beta-mediated phosphorylation in 
            control of epithelial-mesenchymal transition.
  JOURNAL   Nat Cell Biol. 2004 Oct . 6(10):931-40.
  PUBMED    15448698
REFERENCE   2  (bases 1 to 5563)
  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: "Nat Cell 
            Biol. 2004 Oct . 6(10):931-40."
COMMENT     Sequence Label: GFP Snail WT
FEATURES             Location/Qualifiers
     source          1..5563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        129..320
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     277..297
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(322..426)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      453..908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(540..559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     750..771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    915..1036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(926..945)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     980..999
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2058..2768,2769..2771,2772..2774))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2058..2768,2769..2771,2772..2774))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2100..2121)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2449..2468
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2708..2729
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2771..2780
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2819..3022)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2848..2868)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3023..3326
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3501..4089)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3581..3600)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4418..4465
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4490..4509
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4697..5491)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(4697..5491)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4808..4827)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5418..5437
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5526..5563,1..320))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5540..5563,1..112)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
       61 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
      121 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      181 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
      241 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
      301 gctggttctt tccgcctcag gactcttcct ttttcaatat tattgaagca tttatcaggg
      361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
      421 tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc
      481 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      541 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      601 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
      661 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
      721 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
      781 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
      841 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
      901 tacaatttac gcgttaagat acattgatga gtttggacaa accacaacta gaatgcagtg
      961 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1021 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1081 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
     1141 tcagttatct agatccggtg gatcccgggc ccgcggtacc gtcgactgca gaattcgccc
     1201 ttgtactcga gtcagcgggg acatcctgag cagccggact cttggtgctt gtggagcagg
     1261 gacattcggg agaaggtccg agcacacgcc tggcactggt acttcttgac atctgagtgg
     1321 gtctggaggt gggcccgcag gttggagcgg tcagcgaagg cacggctgca gtggggacag
     1381 gagaagggct tctcgccagt gtgggtccgg acatggcctt gtagcagcca gggcctagag
     1441 aaggccttcc cgcaggttcc gcagacgcag ggcagcgtgt ggcttcggat gtgcatcttg
     1501 agggcaccca ggctgaggta ttccttgttg cagtatttgc agttgaaggc ctttcgagcc
     1561 tggagatcct tggcctcaga gagctgggcc agctgcttgg gcacttggcc caagcctggg
     1621 aaggcagcat aggcctcggc ctccaaggaa gagactgaag tagaggagaa ggacgaagga
     1681 gccggtgagg gtgggctggg gggctgggag cctttcccac tgtcctcatc tgacagggag
     1741 gtcagctctg ccaccctggg actctcctgg agccgaaggg aggcccaggc aattggctgg
     1801 gcttggggcg ccaggacaga gtcccagatg agcattggca gcgaggcggt ggggttgagg
     1861 atctccggag gtgggatggc tgccagcagg tgggcctggt cgtagggctg ctggaaggta
     1921 aactctggat tagagtcctg cagctcgctg tagttaggct tccgattggg gtcggagggc
     1981 ttcctgacga ggaaagagcg cggggatcct acaagggcga attcgaagct tgagctcgag
     2041 atctgagtcc ggccggactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     2101 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     2161 tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     2221 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag
     2281 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag
     2341 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc
     2401 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg
     2461 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg
     2521 aagaagtcgt gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc
     2581 agggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc
     2641 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg
     2701 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg
     2761 cccttgctca ccatggtggc gaccggtagc gctagcggat ctgacggttc actaaaccag
     2821 ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg
     2881 agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga
     2941 cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga
     3001 tgtactgcca aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc
     3061 aagtaggaaa gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt
     3121 cattgacgtc aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg
     3181 cagtttaccg taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta
     3241 tgggaacata cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg
     3301 ggccatttac cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc
     3361 ccgtaattga ttactattaa taactaatgc atggcggtaa tacggttatc cacagaatca
     3421 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3481 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3541 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3601 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3661 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3721 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3781 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3841 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3901 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3961 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4021 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4081 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4141 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4201 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga ggctatggca
     4261 gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact tggggggtgg
     4321 ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg tggggtatcg
     4381 acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc aacaccgtgc
     4441 gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc
     4501 cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag atccccgcgc
     4561 tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag
     4621 gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg
     4681 aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     4741 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     4801 cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc acacccagcc
     4861 ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     4921 catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg agcctggcga
     4981 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     5041 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     5101 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     5161 cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     5221 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     5281 ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg gacaggtcgg
     5341 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     5401 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     5461 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     5521 cgatctttgc aaaagcctag gcctccaaaa aagcctcctc act
//
