LOCUS       sequence-8529-ex        6347 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 6347)
  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 6347)
  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: ZP59 23f17
FEATURES             Location/Qualifiers
     source          1..6347
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    31..47
                     /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     36..58
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     55..71
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     55..71
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     90..109
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        90..108
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2463..2481)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2464..2481)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2489..2506)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2489..2505)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2498..2520)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2647..3102
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2734..2753)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2944..2965
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3129..3233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3234..3302,3303..4094)
                     /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(3452..3471)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4265..4853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4754..4773
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5007..5024
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5131..5208
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5209..6291
                     /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(5228..5247)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    6304..6325
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6340..6345,6346..6347,1..16,17..23)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
       61 acagctatga ccatgattac gccaagctct aatacgactc actataggga aagctggtac
      121 gcctgcaggt accggtccgg aattcccggg tcgacgccta accagatcag cccataacat
      181 ggggtctgcc ccatgttatg tatatttcaa agtcaggaat gcatcttttt catcttggat
      241 ttatctttga taatttgatg ttttgtattg atcatttatg aattgactga ttgaattggc
      301 ataatacatt tacctgacta ataaattgta atgtttgacc atattttgct cactctgtaa
      361 ttattaattc aagtagatta cgctgtatcc ttgtttccgc acgtcttgcg tcaggtcagt
      421 agacggacta aaatccaatt gagatggagc attggcacac taattgtgtt ttagcgatcc
      481 ggctgactct tgatattgat tcatgtgtac ttaggtggaa agggcgataa cttccgtcta
      541 ggtcaacaag gtgttgcacg actaacctag attgggtacc cgacctttgt ttgaaggtaa
      601 tattattcta aattaagttc atatgggaaa ccatgggctt ggtgaaaacc aaagttacta
      661 tagagtccag tagtcgggta catttatatt aatgccctaa cctctaatta gaaatgatga
      721 ttgtcttaac tcaaaggtgt acatctaagc gggactaact aaagtatttt agaacgagcg
      781 cgctggtagc ggccatctaa agtattagtc aatttacctc tgtttaatat tcaagactga
      841 ccggagtgtg accgtgaggg acgccttcgg ccgaggcgat ttctctgagc gacatgagca
      901 cccgaatgaa cggatgtaat agttactagt gaagacaaaa ggttcaaaca tttatctaaa
      961 ttacatattg atataatctt cttaatgttt tataattgga gtcagataaa acgaggataa
     1021 atatattaat attctaacca ttctcatatt aaaattggag tcagatatga gttaagttta
     1081 atataaatca acattgtgca ctggaaagac cgaacatgca gtgaaccttc atccaccatt
     1141 ggataccgtc attggaccaa ctgggtggac cttacaaaat ggtgacccat ctccgtgatc
     1201 tcccaactca ggcgagttac gtctttccag cacaaaatgg atttatatca accaagaggt
     1261 ttggatcttc tattctatct tcagctgctg tggtaagtca attcttcttt gcctattaga
     1321 aatgttgagg gaaagctgga cgcaccctnt ataaacacat tgatagaatg agtcggaaat
     1381 gctcaaaaat gttagaccgg aatgcgttat ccggtggggt tgagaattag acaaaaccac
     1441 tttgattatc agggacatga tagcagcagt ataganaaan gccatccctt gctctgaaat
     1501 atctctttgt ctgctaggca gcaagagttg aagcttaaaa ttggaataac atcatccaaa
     1561 ttatattctg gnactcagtg gagagtcaca aattagatcc taaaagtttg aatcaaaaat
     1621 atccattgtt tttaaatcta ctgttttata aaaaagagaa aaatggaaaa aaggaaaaca
     1681 ggaaaaaatg gtttaagatt taaaaacaat tgtaaacttt taaattggat taataacaac
     1741 agtctgttta aaggcatttg catgtgaaaa ataaaggtga gcagaacaga atgatttgat
     1801 tgcaaaaaag acaaataagc tctaatgaaa actctgcttt ttacaaatca gaaatttgcc
     1861 atttaaattt aatggggata aattcaaata tgatgctgat aaattaaaat atccacaata
     1921 ctttaattta atctcgaaac agactaaata taaaagattt tagctttaaa taagccctta
     1981 gacctcctga tggctcaaac cttttcatat tcaaatggcc tgacagagag caaagaagtc
     2041 tcacctcagt gataataaga tgctcagaat gtgtttgctg taattgcata aaacaatgca
     2101 ctgtgttcta aaacactctc cagagaccag atataattat atttaataca tactatttta
     2161 atataatata attacattag gaataaacaa taatgtacat tatattataa tcgccaatca
     2221 tggcactaaa gactagagat gaaacattca ttttgaggag gtttgctttc tttatgctta
     2281 atgaaaacac tgacagaact actatagaat atctgattgt aaaccaatat aatcctcgct
     2341 gaaaaaacac cacctacaat taaaatttga tcagagagtt aaactcacca aaaaaaaaaa
     2401 aaaagggcgg ccgctctaga ggatccaagc ttacgtacgc gtgcatgcga cgtcatagct
     2461 cttctatagt gtcacctaaa ttcaattcac tggccgtcgt tttacaacgt cgtgactggg
     2521 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2581 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2641 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2701 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2761 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2821 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2881 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2941 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3001 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3061 aatttaacgc gaattttaac aaaatattaa cgtttacaat ttcaggtggc acttttcggg
     3121 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     3181 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     3241 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     3301 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     3361 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     3421 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     3481 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     3541 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     3601 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     3661 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     3721 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     3781 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3841 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3901 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3961 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     4021 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     4081 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     4141 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4201 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4261 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4321 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4381 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4441 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4501 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4561 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4621 cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg
     4681 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4741 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4801 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4861 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4921 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4981 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     5041 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcaga gcttgcaatt
     5101 cgcgcgcgaa ggcgaagcgg catttacgtt gacaccatcg aatggcgcaa aacctttcgc
     5161 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     5221 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     5281 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     5341 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     5401 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     5461 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     5521 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gggtcagtgg gctgatcatt
     5581 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     5641 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     5701 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     5761 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     5821 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     5881 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     5941 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     6001 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     6061 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     6121 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     6181 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     6241 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6301 aattaatgtg agttagctca ctcattaggc accccaggct ttacact
//
