LOCUS       sequence-223612-        5567 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 5567)
  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 5567)
  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: Tag5Amyc-GSK3b CA
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        554..572
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      651..660
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1474..1485
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1920..1949
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2002..2021)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2003..2021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2295..2416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2332..2351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2386..2405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2423..2878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2560..2581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2772..2791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2905..3009
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3011..3368
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3011..3202
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3034..3054)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3219..3354
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3403..4197
                     /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             3403..4197
                     /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(3457..3476)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4067..4086
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4385..4404)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4429..4476
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4805..5393
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5294..5313
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt
      541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg
      601 gcggccgctc tagcccgggc ggatcccccg ggctgcagga attcgatatc gccaccatgg
      661 ccggccggcc cagaaccacc gcctttgcgg agagctgcaa gccggtgcag cagccttcag
      721 cttttggcag catgaaagtt agcagagaca aggacggcag caaggtgaca acagtggtgg
      781 caactcctgg gcagggtcca gacaggccac aagaagtcag ctatacagac actaaagtga
      841 ttggaaatgg atcatttggt gtggtatatc aagccaaact ttgtgattca ggagaactgg
      901 tcgccatcaa gaaagtattg caggacaaga gatttaagaa tcgagagctc cagatcatga
      961 gaaagctaga tcactgtaac atagtccgat tgcgttattt cttctactcc agtggtgaga
     1021 agaaagatga ggtctatctt aatctggtgc tggactatgt tccggaaaca gtatacagag
     1081 ttgccagaca ctatagtcga gccaaacaga cgctccctgt gatttatgtc aagttgtata
     1141 tgtatcagct gttccgaagt ttagcctata tccattcctt tggaatctgc catcgggata
     1201 ttaaaccgca gaacctcttg ttggatcctg atactgctgt attaaaactc tgtgactttg
     1261 gaagtgcaaa gcagctggtc cgaggagaac ccaatgtttc gtatatctgt tctcggtact
     1321 atagggcacc agagttgatc tttggagcca ctgattatac ctctagtata gatgtatggt
     1381 ctgctggctg tgtgttggct gagctgttac taggacaacc aatatttcca ggggatagtg
     1441 gtgtggatca gttggtagaa ataatcaagg tcctgggaac tccaacaagg gagcaaatca
     1501 gagaaatgaa cccaaactac acagaattta aattccctca aattaaggca catccttgga
     1561 ctaaggtctt ccgaccccga actccaccgg aggcaattgc actgtgtagc cgtctgctgg
     1621 agtatacacc aactgcccga ctaacaccac tggaagcttg tgcacattca ttttttgatg
     1681 aattacggga cccaaatgtc aaactaccaa atgggcgaga cacacctgca ctcttcaact
     1741 tcaccactca agaactgtca agtaatccac ctctggctac catccttatt cctcctcatg
     1801 ctcggattca agcagctgct tcaaccccca caaatgccac agcagcgtca gatgctaata
     1861 ctggagaccg tggacagacc aataatgctg cttctgcatc agcttccaac tccaccctcg
     1921 agcagaaact catctctgaa gaggatctgt agggcccggt accttaatta attaaggtac
     1981 caggtaagtg tacccaattc gccctatagt gagtcgtatt acaattcact cgatcgccct
     2041 tcccaacagt tgcgcagcct gaatggcgaa tggagatcca atttttaagt gtataatgtg
     2101 ttaaactact gattctaatt gtttgtgtat tttagattca cagtcccaag gctcatttca
     2161 ggcccctcag tcctcacagt ctgttcatga tcataatcag ccataccaca tttgtagagg
     2221 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2281 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2341 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     2401 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     2461 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2521 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2581 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2641 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2701 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2761 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2821 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2881 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2941 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3001 aggaagaatc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3061 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3121 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3181 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3241 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3301 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3361 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     3421 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3481 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3541 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3601 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3661 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3721 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3781 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3841 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagaacatca ggggctcgcg
     3901 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     3961 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4021 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4081 gatattgctg aagaacttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4141 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4201 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4261 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4321 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4381 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     4441 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4501 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4561 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4621 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4681 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4741 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4801 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4861 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4921 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4981 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5041 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5101 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5161 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5221 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5281 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5341 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     5401 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     5461 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     5521 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
