LOCUS       Exported                5563 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    GFP Snail WT
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5563
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     regulatory      614..623
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             620..1336
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             620..1336
                     /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(665..686)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(926..945)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1273..1294
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2358..2479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2395..2414)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2449..2468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2486..2941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2623..2644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2835..2854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2968..3072
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3074..3431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3097..3117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3282..3417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3344..3363
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3466..4260
                     /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(3520..3539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4130..4149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4448..4467)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4492..4539
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4868..5456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5357..5376
                     /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 agcgctaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      661 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      721 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      781 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      841 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      901 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      961 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1021 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1081 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1141 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1201 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1261 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1321 ggacgagctg tacaagtccg gccggactca gatctcgagc tcaagcttcg aattcgccct
     1381 tgtaggatcc ccgcgctctt tcctcgtcag gaagccctcc gaccccaatc ggaagcctaa
     1441 ctacagcgag ctgcaggact ctaatccaga gtttaccttc cagcagccct acgaccaggc
     1501 ccacctgctg gcagccatcc cacctccgga gatcctcaac cccaccgcct cgctgccaat
     1561 gctcatctgg gactctgtcc tggcgcccca agcccagcca attgcctggg cctcccttcg
     1621 gctccaggag agtcccaggg tggcagagct gacctccctg tcagatgagg acagtgggaa
     1681 aggctcccag ccccccagcc caccctcacc ggctccttcg tccttctcct ctacttcagt
     1741 ctcttccttg gaggccgagg cctatgctgc cttcccaggc ttgggccaag tgcccaagca
     1801 gctggcccag ctctctgagg ccaaggatct ccaggctcga aaggccttca actgcaaata
     1861 ctgcaacaag gaatacctca gcctgggtgc cctcaagatg cacatccgaa gccacacgct
     1921 gccctgcgtc tgcggaacct gcgggaaggc cttctctagg ccctggctgc tacaaggcca
     1981 tgtccggacc cacactggcg agaagccctt ctcctgtccc cactgcagcc gtgccttcgc
     2041 tgaccgctcc aacctgcggg cccacctcca gacccactca gatgtcaaga agtaccagtg
     2101 ccaggcgtgt gctcggacct tctcccgaat gtccctgctc cacaagcacc aagagtccgg
     2161 ctgctcagga tgtccccgct gactcgagta caagggcgaa ttctgcagtc gacggtaccg
     2221 cgggcccggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     2281 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2341 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2401 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2461 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2521 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2581 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2641 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2701 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2761 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2821 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2881 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2941 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     3001 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     3061 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     3121 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3181 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3241 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3301 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3361 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3421 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3481 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3541 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3601 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3661 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3721 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3781 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3841 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3901 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3961 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     4021 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4081 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4141 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4201 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4261 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4321 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4381 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4441 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4501 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4561 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4621 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4681 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4741 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4801 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4861 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4921 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4981 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     5041 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5101 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5161 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5221 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5281 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5341 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5401 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5461 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5521 ttcctgcgtt atcccctgat tctgtggata accgtattac cgc
//
