LOCUS       sequence-137677-        6813 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 6813)
  AUTHORS   Miller JR, Rowning BA, Larabell CA, Yang-Snyder JA, Bates RL, Moon 
            RT
  TITLE     Establishment of the dorsal-ventral axis in Xenopus embryos 
            coincides with the dorsal enrichment of dishevelled that is 
            dependent on cortical rotation.
  JOURNAL   J Cell Biol. 1999 Jul 26;146(2):427-37. doi: 10.1083/jcb.146.2.427.
  PUBMED    10427095
REFERENCE   2  (bases 1 to 6813)
  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: "10.1083/jcb.146.2";
            journalName: "J Cell Biol"; date: "1999-07-26- 26"; volume: "146"; 
            issue: "2"; pages: "427-37"
COMMENT     Sequence Label: XE124 XDsh delta DEP-GFP-CS2+
FEATURES             Location/Qualifiers
     source          1..6813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        16..34
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     16..33
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2079..2792
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2109..2137)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2732..2752
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    2846..2980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2870..2889)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2924..2943
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3093..3113)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3093..3111)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3132..3148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3132..3148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3145..3167)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3156..3172
                     /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(3180..3186,3187..3204,3205..3210))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3225..3246
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3363..3380)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3534..4122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3614..3633)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4293..5084,5085..5153))
                     /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     4916..4935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5154..5258)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5284..5739)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5421..5442)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5633..5652
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5810..6794
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gtcggagcaa gcttgattta ggtgacacta tagaatacaa gctacttgtt ctttttgcag
       61 gatcaaccat ggcggagact aaagtgattt accatctgga tgaggaggag accccgtacc
      121 tggtgaaggt ccctgtcccg gccactgaca tccgcctgag ggacttcaag gctgcgctgg
      181 gccgaggcca cgccaaatat ttcttcaagg ccatggacca agacttcggg gtagttaagg
      241 aggagatctc agatgacaat gccaagctgc cgtgctttaa tggccgtgtg gtgtcgtggc
      301 ttgtgtcctc cgagggttct cagccggatt ctgcaccccc ggcccctgct acagaggtca
      361 gacctgaacc cccccctcct gttccacccc cgattccacc accccctgca gagagaacca
      421 gcggtattgg agattcccgg cctccctcct ttcatcccaa cgtgtcgggc agtactgagc
      481 agctggatca ggacaacgag tctgtcatct ccatgcgcag agaccgagtc aggcggagag
      541 agagcagtga gcaagcaggt gttggcagag gggtgaatgg acggacagag cggcacctat
      601 ctgggtatga atcttcctcg actctattga ccagtgagat tgagacgagt atctgtgatt
      661 cggaggagga cgacaccatg agcaggttca gcagttccac ggagcagagc agtgcctcac
      721 gccttctcaa acggcaccgg cgccgccgca agcagagacc cccgcgcctg gaacggacgt
      781 cctcattcag cagcgtcacc gactccacca tgtccctaaa catcatcact gtgacgctga
      841 atatggagaa gtacaatttc ctgggcatct ccatcgtggg gcagagtaac gagcggggag
      901 acgggggcat ttacattggc tctataatga agggcggagc tgtggcagct gacggcagaa
      961 ttgaacccgg ggacatgttg ctacaggtga atgacattaa ctttgagaat atgagcaatg
     1021 acgatgcagt ccgtgtgctc agggatattg tccataaacc ggggcccatt gtcctgaccg
     1081 tggccaaatg ctgggaccct tcccctcagg ggtactttac cctcccccgc aatgagccca
     1141 tccaccctat tgatccggct gcctgggtct ctcactcggc tgccctgagc gggtccttcc
     1201 cagtgtaccc cgggagcgcc tccatgagca gcatgacctc cagcacctct gtcactgaga
     1261 ctgagctttc ccatgcgctc cctccggtgt cactcttcag cctctccgta cacaccgatt
     1321 tggcttctgt tgtgaaggtc atggcttccc ctgagtccgg cctcgagctg actggctgtg
     1381 agaactatat ggccaacctc tctctgaatg acaacgatgg atccagcggc gcttccgacc
     1441 aggacactct ggcccctctt cccctccctg gggcttcccc atggcctcta ctccccacct
     1501 tctcttacca gtatcaagcc ccacacccgt acagcactca gccacctgcc taccatgagc
     1561 tctcctcgta cagctacggc atgggcagtg cgggcagcca gcacagcgag ggcagcagaa
     1621 gcagtggatc caaccgcagc gatgggggtc ggggcatgca gaaggatgac cgttccggag
     1681 tagcgggagt gggtggggga gactcgaagt cgggatctgg tagtgagtct gagtattcca
     1741 cccgcagcag cataaggaga gtaggtggtg gggaggctgg tccccccagt gagcgctcca
     1801 cttcatcccg acttccacct caccatccac cctctgttca ctcctatgct gctcctggtg
     1861 tccctctctc ctacaatcca atgatgctca tgatgatgcc gccccctcca ttacctccac
     1921 ctggggtttg tcctcccaat tcttctgtac cacctggagc tccacctctt gtacgggatc
     1981 ttgcgtctgt tcccccggag ctgactgcca gccgccagtc tttccatatg gccatgggca
     2041 accctagcga gttctttgtg gatgtcatgc atcgattgat gagtaaagga gaagaacttt
     2101 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
     2161 ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt
     2221 gcactactgg aaaactacct gttccatggc caacacttgt cactactctc acttatggtg
     2281 ttcaatgctt ttcaagatac ccagatcata tgaaacggca tgactttttc aagagtgcca
     2341 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     2401 cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta
     2461 ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac tataactcac
     2521 acaatgtata catcatggca gacaaacaaa agaatggaat caaagttaac ttcaaaatta
     2581 gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa
     2641 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt
     2701 cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg
     2761 ggattacaca tggcatggat gaactataca aataagaatt caaggcctct cgagcctcta
     2821 gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg
     2881 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2941 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     3001 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc
     3061 aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     3121 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3181 aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3241 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3301 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3361 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3421 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3481 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3541 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3601 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3661 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3721 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3781 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3841 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3901 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3961 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4021 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4081 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4141 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4201 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4261 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4321 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4381 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4441 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4501 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4561 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4621 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4681 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4741 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4801 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4861 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4921 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4981 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5041 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5101 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5161 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5221 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5281 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5341 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5401 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5461 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5521 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5581 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5641 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5701 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     5761 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca
     5821 attcaatatg gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca
     5881 attcaatatg gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca
     5941 attcaatatg gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca
     6001 actggggagg ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc
     6061 aagtccgcca tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca
     6121 ggatcaatat ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt
     6181 caatactatg tattggccct atgccatata gtattccata tatgggtttt cctattgacg
     6241 tagatagccc ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc
     6301 catagccact cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca
     6361 tatgggcggt cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg
     6421 gcccgcctgg ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg
     6481 atggctcatt gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa
     6541 tggcccactt ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt
     6601 ggaagtacgc cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc
     6661 gcgatggctg ccaagtacat ccccattgac gtcaatgggg aggggcaatg acgcaaatgg
     6721 gcgttccatt gacgtaaatg ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg
     6781 ctcgtttagg gaaccgccat tctgcctggg gac
//
