LOCUS       sequence-154381-        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 KD
FEATURES             Location/Qualifiers
     source          1..5567
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        621..639
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             1541..1552
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1987..2016
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2069..2088)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2070..2088)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2362..2483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2399..2418)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2453..2472
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2490..2945)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2627..2648)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2839..2858
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2972..3076
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3078..3435
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3078..3269
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3101..3121)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3286..3421
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3470..4264
                     /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             3470..4264
                     /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(3524..3543)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4134..4153
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4452..4471)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4496..4543
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4872..5460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5361..5380
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgattacg ccaagctcga aattaaccct cactaaaggg aacaaaagct ggagctccac
      661 cgcggtggcg gccgctctag cccgggcgga tcccccgggc tgcaggaatt cgatatcgcc
      721 accatgtcag ggcggcccag aaccacctcc tttgcggaga gctgcaagcc ggtgcagcag
      781 ccttcagctt ttggcagcat gaaagttagc agagacaagg acggcagcaa ggtgacaaca
      841 gtggtggcaa ctcctgggca gggtccagac aggccacaag aagtcagcta tacagacact
      901 aaagtgattg gaaatggatc atttggtgtg gtatatcaag ccaaactttg tgattcagga
      961 gaactggtcg ccatcgcgaa agtattgcag gacaagagat ttaagaatcg agagctccag
     1021 atcatgagaa agctagatca ctgtaacata gtccgattgc gttatttctt ctactccagt
     1081 ggtgagaaga aagatgaggt ctatcttaat ctggtgctgg actatgttcc ggaaacagta
     1141 tacagagttg ccagacacta tagtcgagcc aaacagacgc tccctgtgat ttatgtcaag
     1201 ttgtatatgt atcagctgtt ccgaagttta gcctatatcc attcctttgg aatctgccat
     1261 cgggatatta aaccgcagaa cctcttgttg gatcctgata ctgctgtatt aaaactctgt
     1321 gactttggaa gtgcaaagca gctggtccga ggagaaccca atgtttcgta tatctgttct
     1381 cggtactata gggcaccaga gttgatcttt ggagccactg attatacctc tagtatagat
     1441 gtatggtctg ctggctgtgt gttggctgag ctgttactag gacaaccaat atttccaggg
     1501 gatagtggtg tggatcagtt ggtagaaata atcaaggtcc tgggaactcc aacaagggag
     1561 caaatcagag aaatgaaccc aaactacaca gaatttaaat tccctcaaat taaggcacat
     1621 ccttggacta aggtcttccg accccgaact ccaccggagg caattgcact gtgtagccgt
     1681 ctgctggagt atacaccaac tgcccgacta acaccactgg aagcttgtgc acattcattt
     1741 tttgatgaat tacgggaccc aaatgtcaaa ctaccaaatg ggcgagacac acctgcactc
     1801 ttcaacttca ccactcaaga actgtcaagt aatccacctc tggctaccat ccttattcct
     1861 cctcatgctc ggattcaagc agctgcttca acccccacaa atgccacagc agcgtcagat
     1921 gctaatactg gagaccgtgg acagaccaat aatgctgctt ctgcatcagc ttccaactcc
     1981 accctcgagc agaaactcat ctctgaagag gatctgtagg gcccggtacc ttaattaatt
     2041 aaggtaccag gtaagtgtac ccaattcgcc ctatagtgag tcgtattaca attcactcga
     2101 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg agatccaatt tttaagtgta
     2161 taatgtgtta aactactgat tctaattgtt tgtgtatttt agattcacag tcccaaggct
     2221 catttcaggc ccctcagtcc tcacagtctg ttcatgatca taatcagcca taccacattt
     2281 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2341 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2401 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2461 tccaaactca tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt
     2521 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2581 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2641 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2701 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2761 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2821 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2881 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2941 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3001 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3061 ttgaaaaagg aagaatcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3121 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3181 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3241 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3301 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3361 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3421 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3481 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3541 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3601 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3661 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3721 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3781 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3841 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3901 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag aacatcaggg
     3961 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4021 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4081 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4141 tacccgtgat attgctgaag aacttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4201 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4261 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4321 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4381 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4441 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4501 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4561 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4621 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4681 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4741 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4801 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4861 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4921 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4981 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5041 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5101 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5161 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5221 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5281 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5341 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5401 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5461 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5521 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgc
//
