LOCUS       sequence-137675-        6546 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 6546)
  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 6546)
  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: XE122 XDsh delta DIX-GFP-CS2+
FEATURES             Location/Qualifiers
     source          1..6546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..41
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     23..40
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1819..2532
                     /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
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1849..1877)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2472..2492
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    2586..2720
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2610..2629)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2664..2683
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2833..2853)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2833..2851)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2872..2888)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2872..2888)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2885..2907)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2896..2912
                     /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(2920..2926,2927..2944,2945..2950))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2965..2986
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3103..3120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3274..3862)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3354..3373)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4033..4824,4825..4893))
                     /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     4656..4675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4894..4998)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5024..5479)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5161..5182)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5373..5392
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5550..6534
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 tggggacgtc ggagcaagct tgatttaggt gacactatag aatacaagct acttgttctt
       61 tttgcaggat ctaccatgag tgagcaagca ggtgttggca gaggggtgaa tggacggaca
      121 gagcggcacc tatctgggta tgaatcttcc tcgactctat tgaccagtga gattgagacg
      181 agtatctgtg attcggagga ggacgacacc atgagcaggt tcagcagttc cacggagcag
      241 agcagtgcct cacgccttct caaacggcac cggcgccgcc gcaagcagag acccccgcgc
      301 ctggaacgga cgtcctcatt cagcagcgtc accgactcca ccatgtccct aaacatcatc
      361 actgtgacgc tgaatatgga gaagtacaat ttcctgggca tctccatcgt ggggcagagt
      421 aacgagcggg gagacggggg catttacatt ggctctataa tgaagggcgg agctgtggca
      481 gctgacggca gaattgaacc cggggacatg ttgctacagg tgaatgacat taactttgag
      541 aatatgagca atgacgatgc agtccgtgtg ctcagggata ttgtccataa accggggccc
      601 attgtcctga ccgtggccaa atgctgggac ccttcccctc aggggtactt taccctcccc
      661 cgcaatgagc ccatccaccc tattgatccg gctgcctggg tctctcactc ggctgccctg
      721 agcgggtcct tcccagtgta ccccgggagc gcctccatga gcagcatgac ctccagcacc
      781 tctgtcactg agactgagct ttcccatgcg ctccctccgg tgtcactctt cagcctctcc
      841 gtacacaccg atttggcttc tgttgtgaag gtcatggctt cccctgagtc cggcctcgag
      901 gtccgagatc gcatgtggct caaaatcacc attcccaacg ccttcctggg ttctgatgtg
      961 gtggactggc tctatcacca cgtggaggga ttccaggacc ggcgagaggc tcgcaagttt
     1021 gccagtaacc tgctgaaggc cggcttcatt cgccacacag tgaacaagat cacattctcc
     1081 gagcagtgtt actatatctt tggggatctg actggctgtg agaactatat ggccaacctc
     1141 tctctgaatg acaacgatgg atccagcggc gcttccgacc aggacactct ggcccctctt
     1201 cccctccctg gggcttcccc atggcctcta ctccccacct tctcttacca gtatcaagcc
     1261 ccacacccgt acagcactca gccacctgcc taccatgagc tctcctcgta cagctacggc
     1321 atgggcagtg cgggcagcca gcacagcgag ggcagcagaa gcagtggatc caaccgcagc
     1381 gatgggggtc ggggcatgca gaaggatgac cgttccggag tagcgggagt gggtggggga
     1441 gactcgaagt cgggatctgg tagtgagtct gagtattcca cccgcagcag cataaggaga
     1501 gtaggtggtg gggaggctgg tccccccagt gagcgctcca cttcatcccg acttccacct
     1561 caccatccac cctctgttca ctcctatgct gctcctggtg tccctctctc ctacaatcca
     1621 atgatgctca tgatgatgcc gccccctcca ttacctccac ctggggtttg tcctcccaat
     1681 tcttctgtac cacctggagc tccacctctt gtacgggatc ttgcgtctgt tcccccggag
     1741 ctgactgcca gccgccagtc tttccatatg gccatgggca accctagcga gttctttgtg
     1801 gatgtcatgc atcgattgat gagtaaagga gaagaacttt tcactggagt tgtcccaatt
     1861 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
     1921 ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct
     1981 gttccatggc caacacttgt cactactttc acttatggtg ttcaatgctt ttcaagatac
     2041 ccagatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     2101 gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt
     2161 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
     2221 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca
     2281 gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga
     2341 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     2401 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     2461 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
     2521 gaactataca aataagaatt caaggcctct cgagcctcta gaactatagt gagtcgtatt
     2581 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     2641 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     2701 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     2761 gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc
     2821 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     2881 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat
     2941 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3001 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3061 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3121 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3181 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3241 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3301 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3361 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3421 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3481 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3541 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3601 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3661 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3721 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3781 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3841 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3901 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4021 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4141 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4201 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4261 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4321 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4381 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4441 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4501 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4561 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4621 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4681 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4741 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4801 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4861 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4921 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4981 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     5041 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5101 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5161 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5221 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     5281 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     5341 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5401 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5461 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5521 tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg
     5581 actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg
     5641 actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg
     5701 caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg
     5761 gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg
     5821 gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata
     5881 tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg tattggccct
     5941 atgccatata gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg
     6001 gcggtcccat ataccatata tggggcttcc taataccgcc catagccact cccccattga
     6061 cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg
     6121 tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc
     6181 attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat
     6241 atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat
     6301 caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat
     6361 tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat
     6421 ccccattgac gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg
     6481 ggcggtaggc gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat
     6541 tctgcc
//
