LOCUS       sequence-245429-        7139 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 7139)
  AUTHORS   Liu JA, Wu MH, Yan CH, Chau BK, So H, Ng A, Chan A, Cheah KS, 
            Briscoe J, Cheung M
  TITLE     Phosphorylation of Sox9 is required for neural crest delamination 
            and is regulated downstream of BMP and canonical Wnt signaling.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Feb 19;110(8):2882-7. doi: 
            10.1073/pnas.1211747110. Epub 2013 Feb 4.
  PUBMED    23382206
REFERENCE   2  (bases 1 to 7139)
  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: 
            "10.1073/pnas.1211747110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-02-19- 19"; volume: "110"; issue: "8"; pages: "2882-7"
COMMENT     Sequence Label: pCIG-V5-Snail2-IRES-nls-GFP
FEATURES             Location/Qualifiers
     source          1..7139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..659
                     /label=chicken beta-actin promoter
     intron          660..1676
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1600..1622
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1684..1703
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1743..1784
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2659..3211
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2803..2820)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3030..3049
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3238..3258
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3262..3282
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3286..3306
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(3316..3318,3319..3321,3322..4035)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3316..3318,3319..3321,3322..4032)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3361..3382)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3622..3641)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3969..3990
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4127..4146)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4192..4247
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4246..4265)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4638..4654)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4638..4654)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4651..4673)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4662..4678
                     /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(4686..4692,4693..4710,4711..4716))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4731..4752
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4811..5006
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4857..4992
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5012..5146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5049..5068)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5103..5122
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5214..5231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5385..5973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5465..5484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6144..6935,6936..7004))
                     /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     6767..6786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7005..7109)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      541 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct
      961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg
     1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt
     1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc
     1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg
     1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg
     1261 ggccgcctcg ggccggggag ggctcggggg aggggcgcgg cggcccccgg agcgccggcg
     1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc
     1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc
     1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg
     1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc
     1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga
     1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc
     1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg ctcgaggcca
     1741 tgggtaagcc tatccctaac cctctcctcg gtctcgattc tacgatgcca cgctccttcc
     1801 tggtcaagaa acatttcaat tcatccaaga agccaaacta cagcgaactg gacactcata
     1861 cagtgattat atccccatac ctgtatgaaa gctacccagt ccctatcata ccgcagccag
     1921 agatcctgag ctctgtagct tacaacccca ttactgtgtg gactacaacc gggctgctac
     1981 cgtctcctct acccaatgac ctctctccgc tttctggata cccctcatct ttgggaaggg
     2041 tcagcccacc tccaccatcc gacacctcct ccaaagatca cagcggttca gaaagtccca
     2101 ttagcgatga agaagagaga atccagtcca agctttcaga ccctcatgcc atcgaagctg
     2161 aaaagtttca gtgcggttta tgcaacaaga cctattccac tttctctggg ttggccaaac
     2221 acaagcagct gcactgtgat gcccagtcta ggaaatcgtt cagctgcaag tactgtgaca
     2281 aggagtatgt cagcctggga gcacttaaga tgcacatcag gacccacacg ctaccttgcg
     2341 tctgcaagat ctgcggcaag gctttctcca gaccctggct acttcaaggg cacatcagaa
     2401 cccacactgg agagaaaccg ttttcctgtc ctcactgcaa cagggctttc gcagacagat
     2461 ccaatctgag ggctcatctg cagacccact cggatgtgaa gaaataccag tgcaaaaatt
     2521 gctccaaaac tttctccaga atgtctcttc tgcacaaaca tgaggaatct ggctgctgcg
     2581 tagcacactg aatcgataag cttgatatcg aattcatgcc acgctccttc ctggtcaaga
     2641 aacatttcaa ttcatccaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt
     2701 ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac
     2761 ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc
     2821 aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga agacaaacaa
     2881 cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg
     2941 gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg
     3001 tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc
     3061 tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct cggtacacat
     3121 gctttacatg tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg
     3181 tggttttcct ttgaaaaaca cgatgataat atggccacaa ccatggtgca cgtggatcca
     3241 aaaaagaaga gaaaggtaga tccaaaaaag aagagaaagg tagatccaaa aaagaagaga
     3301 aaggtacacg tgtccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3361 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     3421 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     3481 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     3541 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     3601 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3661 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3721 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     3781 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     3841 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3901 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     3961 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     4021 gagctgtaca agtaaagcgg ccgcaattca ctcctcaggt gcaggctgcc tatcagaagg
     4081 tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     4141 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     4201 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4261 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4321 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     4381 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     4441 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     4501 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     4561 tcttatggag atccctcgac ctgcaccgtc gaccagctgg tcgacggtgc accgtcgacc
     4621 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4681 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4741 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4801 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4861 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4921 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     4981 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     5041 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5101 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     5161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5221 actgactcgc tgcgctcggt cgttcagctg cggcgagcgg tatcagctca ctcaaaggcg
     5281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5581 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5821 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtc
//
