LOCUS       sequence-8527-ex        6966 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 6966)
  AUTHORS   Weidinger G, Thorpe CJ, Wuennenberg-Stapleton K, Ngai J, Moon RT
  TITLE     The Sp1-related transcription factors sp5 and sp5-like act 
            downstream of Wnt/beta-catenin signaling in mesoderm and 
            neuroectoderm patterning.
  JOURNAL   Curr Biol. 2005 Mar 29. 15(6):489-500.
  PUBMED    15797017
REFERENCE   2  (bases 1 to 6966)
  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: "Curr Biol. 
            2005 Mar 29. 15(6):489-500."
COMMENT     Sequence Label: ZP62 Cystathionine beta-synthase
FEATURES             Location/Qualifiers
     source          1..6966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    219..240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(255..260,261..278,279..285)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    293..309
                     /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)."
     primer_bind     298..320
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     317..333
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     317..333
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     352..371
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        352..370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3344..3362)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3345..3362)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3370..3387)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3370..3386)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3379..3401)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3528..3983
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3615..3634)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3825..3846
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4010..4114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4115..4183,4184..4975)
                     /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     complement(4333..4352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5146..5734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5635..5654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5888..5905
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        6012..6089
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(6090..6966,1..206)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(6109..6128)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
       61 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
      121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      181 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      241 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      301 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctaatacgac
      361 tcactatagg gaaagctggt acgcctgcag gtaccggtcc ggaattcccg ggtcgaccca
      421 cgcgtccgtc tctgtaatcg gacctccggc tctcaagttt ctctccatct gatagtcagg
      481 gtaaaccatg ccagcagtac catctagttc agaatcagtt ggcactggcc ccgtctgccc
      541 acatgctgcc aaaatgctct caaatgctgc aaatggagag tctggcatca acggggatgc
      601 tgacgatgtc ttcaacggaa aggctggaca caagctgaat gaaaccaacg ggcgccactt
      661 aaagagtctg aacggacagc acatggagaa gggctccgag gtcagcatgg agaggaaatg
      721 gattcgaccg gatttgccca gcagatgcac ctggaagctt ggtgacctga atatggagtc
      781 accccacaac cactaccaac ggacaaagac gtccggtatc ctccccaaca tcttaagcaa
      841 gattggagag accccaatgg ttcgcatgaa caagattccc aaagcttttg gcctcaagtg
      901 tgaactcttg gctaaatgcg agttcttcaa tgctggtggc agtgtgaagg acagaatcag
      961 cttgcgaatg atagaagatg ctgaacgtga tggcaccctc aaacctggag acaccatcat
     1021 tgagcctaca tcaggaaaca caggtatcgg tcttgctctg gcagcagcgg tcaaaggtta
     1081 tcgctgcatt attgtcatgc ctgagaagat gagcatggaa aaggtggatg tgctccgtgc
     1141 tctgggtgct gagattgttc gtaccccaac ctcagcccgc ttcgactctc ctgagtctca
     1201 tgtgggggtc gcgtggcgtt tgaagaatga gattccaaat gctcacattc ttgaccagta
     1261 ccgcaatccc agtaaccccc ttgcccacta tgacaccact gcagaggaga tcctggagca
     1321 gtgtggcggt aaaatagaca tgctggtcgc aggagctggc actggtggca ccatcactgg
     1381 catcgctcgc aagctaaagg agaaatgccc tgatatcaag atcattggag tggatccgga
     1441 gggctctatt ctagctgagc cagacgagtt gaataagaca gataagactc agtatgaggt
     1501 tgaaggcatc gggtatgatt tcatccccac tgttctggac agatctgtgg tggataattg
     1561 gtacaagtcc aatgatgagg agtccttcgc catgtctcgt atgctcatca gagaggaggg
     1621 tcttctgtgc ggtggtagtt ctggcacagc catgtctgca gcagtcaacg tagccaaaga
     1681 gcttaaggaa ggccagcgct gtgttgtcat tctacccgac tccatccgca actacatgtc
     1741 taaattcctc agtgacaagt ggatgtttga aaagggcttc atgagcgagg aggatcttgt
     1801 tgtcaataag ccatggtggt ggaatttgac tcttcaggag cttcgcctct cggctccact
     1861 gacggtgctt cctttggtgt ccatcaagaa gaccattcag atcctcaaag aaaaggcttt
     1921 tgaccaggct cctgtggtgg acgaggcggg gcatattctg gggatggtca cgttggggaa
     1981 catgctcgcg tctgttctgg ctggcaaggt cagaccatcc gatcccataa gcaaagtgct
     2041 ctacaaacag tttaaacagg tgcgtctgac tgataacctt ggcaaactct cacgtatcct
     2101 tgagacggat catttcgccc ttgttgtgca cgaacagatt cagtacatga gtgacggatc
     2161 ccccaagatg aggcagatgg tgttcggggt ggtgacggcc attgacctgc tcaactacgt
     2221 gaccatgcgt gagcgaaggg aacgcacgct gtctgagtgc tcgttgtcag aagatcagtg
     2281 aacaacctga tcacaattct ggagcaaact atttttccat cttaattttg tatttttctt
     2341 ttgtagctta tttctctgca agtggcacgc tccatttcaa gaactatgct ttttatatat
     2401 gagcctgtca gtagctttac atgtcatttg actgatctgt agtctgtgtt cactggatcc
     2461 atttttaaaa aatcttcagt ctctaatatt tagcttttgt tcagtttagg tgctttatta
     2521 gtcatgatga agtaatctta caggtcaatc agctcccatt tgcagatgtg tttacgttgc
     2581 agatttgatt ttgattaagg tgtatatatt tacatgaatc tattttattc tttgaaaaac
     2641 tcagtaggct atgcggtttg gtaggatttg gtttggttta ggtgaagtgc tttattcagg
     2701 tgatctcttg aatgcagcgc cgtctagtgg cagtggcagt ttacagcagg cgtggacact
     2761 ttactttctt caatcatgct cttctaagca caacgatccc tccgctacac gtggagagct
     2821 ttacaggcat gactggtcat gattgacaaa gcaaacaatt cgtgtgtttg ggaaaccaaa
     2881 ctacacccag atggggtgac ttggtatttt tcttgtatta aaacaagaca tgattttgag
     2941 tgtaaataag tgacggtggt ttgaaattac tttaaacgca ccacacttgt gtttaagaaa
     3001 tttgatcacg cttttcattt ttctatgttc ttttggtctt gtaaatcaaa actgtaattt
     3061 atggaggctg aataagattg tgtgtatata agagttcatg tgagtgaatg aatgtttggg
     3121 gttgctctgc atgctgctgg aggtctctgc ctcaacagaa cgtcttattt ttctgttatg
     3181 aatctcatgt cgtttgattt gcttttcatt gaatcggatt gctgtaataa tatgtactag
     3241 ttacaaataa aggctttatt tacttaaaaa aaaaaaaaaa aaaaagggcg gccgctctag
     3301 aggatccaag cttacgtacg cgtgcatgcg acgtcatagc tcttctatag tgtcacctaa
     3361 attcaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     3421 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     3481 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     3541 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3601 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3661 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3721 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3781 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3841 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3901 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3961 caaaatatta acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4021 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4201 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4261 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4321 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4381 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4441 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4501 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4561 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4621 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4681 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4741 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4801 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4861 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4921 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4981 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5041 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5101 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5161 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5221 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5281 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5341 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5401 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5461 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5521 atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5581 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5641 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5701 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5761 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5821 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5881 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     5941 ccccgcgcgt tggccgattc attaatgcag agcttgcaat tcgcgcgcga aggcgaagcg
     6001 gcatttacgt tgacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     6061 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
     6121 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     6181 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     6241 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     6301 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     6361 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     6421 tgcacaatct tctcgcgcaa cgggtcagtg ggctgatcat taactatccg ctggatgacc
     6481 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     6541 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     6601 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     6661 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     6721 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     6781 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     6841 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     6901 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
     6961 aggatt
//
