LOCUS       sequence-42101-e        8737 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 8737)
  AUTHORS   Green JL, Inoue T, Sternberg PW
  TITLE     Opposing Wnt pathways orient cell polarity during organogenesis.
  JOURNAL   Cell. 2008 Aug 22;134(4):646-56.
  PUBMED    18724937
REFERENCE   2  (bases 1 to 8737)
  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: "Cell."; 
            date: "2008-08-22"; volume: "134(4)"; pages: "646-56"
COMMENT     Sequence Label: POPTOP
FEATURES             Location/Qualifiers
     source          1..8737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1698..1718)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1698..1716)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1737..1753)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1737..1753)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1750..1772)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1761..1777
                     /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(1785..1791,1792..1809,1810..1815))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1830..1851
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1968..1985)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2139..2727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2219..2238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2898..3689,3690..3758))
                     /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     3521..3540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3759..3863)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3889..4344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4026..4047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4238..4257
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4471..4493
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4485..4502
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4486..4502
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4512..4531
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4512..4530
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(5446..5457)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    8149..8197
                     /note="synthetic polyadenylation signal"
     misc_feature    8211..8302
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 ctaggatgtc tgattccgct gttgttgccg ccgctgtcga gccaaaggtc ccaaaggcta
       61 aggccgccaa ggccgccaag ccaaccaaag ttgccaaggc caaggctcca gtcgctcacc
      121 caccatacat caacatgatc aaggaggcca tcaagcagct caaggaccgc aaaggagctt
      181 ccaagcaggc aatcctcaag ttcatctccc aaaactacaa gctcggagac aatgtcatcc
      241 agatcaatgc tcatctccgt caggctctca agcgtggtgt caccagcaag gcccttgttc
      301 aagccgccgg atccggagcc aacggacgtt tccgtgtgcc agagaaggcc gccgccgcca
      361 agaagccagc agcagccaag aagccagcag cagccaagaa accagcagcc gccaagaaag
      421 ccactggaga gaagaaggca aagaagccag ccgctgccaa gccaaagaag gccgccaccg
      481 gagataagaa ggtcaagaag gccaagagcc caaagaaggt tgccaagcca gctgccaaga
      541 aggtcgccaa gtctccagcc aagaaggccg ctccaaagaa gatcgccaag ccagctgcca
      601 agaaggccgc caagccagcc gccaaggccg catgcatggt ctcaaagggt gaagaagata
      661 acatggcaat tattaaagag tttatgcgtt tcaaggtgca tatggaggga tctgtcaatg
      721 ggcatgagtt tgaaattgaa ggtgaaggag aaggccgacc atatgaggga acacaaaccg
      781 caaaactaaa ggtaagttta aacatatata tactaactaa ccctgattat ttaaattttc
      841 aggtaactaa aggcggacca ttaccattcg cctgggacat cctctctcca cagttcatgt
      901 atggaagtaa agcttatgtt aaacatccgg cagatatacc agattatttg aaactttcat
      961 tcccggaggg ttttaagtgg gaacgcgtaa tgaattttga agacggagga gttgttacag
     1021 tgacgcaaga ctcaaggtaa gtttaaacag ttcggtacta actaaccata catatttaaa
     1081 ttttcagcct ccaagatgga gaatttattt ataaagtcaa acttcgagga acgaatttcc
     1141 cctcggatgg acctgttatg cagaagaaga ctatgggatg ggaagcttca agtgaaagaa
     1201 tgtaccctga agacggtgct cttaagggag agattaaaca acgtcttaaa ttgaaagatg
     1261 gaggacatta cgatgctgag gtaagtttaa acatgatttt actaactaac taatctgatt
     1321 taaattttca ggtgaagaca acttacaaag ccaaaaaacc agttcagctg ccaggagcgt
     1381 acaatgttaa tattaaactg gatatcacct cccacaacga ggattacact atcgttgagc
     1441 aatatgaaag agctgaaggg cggcactcga caggtggcat ggatgaattg tataagaagc
     1501 tttggaattc gctagccggc catacaagta atccggatga tcgacgccaa cgtcgttgaa
     1561 ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt attctctttg
     1621 tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat gaggggcccg
     1681 gtacccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     1741 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1801 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1861 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     1921 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     1981 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2041 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2101 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2161 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2221 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2281 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2341 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2401 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2461 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2521 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     2581 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2641 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2701 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2761 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2821 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     2881 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     2941 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3001 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3061 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3121 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3181 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3241 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3301 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3361 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3421 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3481 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3541 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3601 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3661 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3721 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3781 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3841 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg
     3901 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3961 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     4021 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     4081 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     4141 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     4201 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     4261 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     4321 ttaatgcgcc gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag
     4381 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     4441 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     4501 gtgagcgcgc gtaatacgac tcactatagg gcgaattgga gctccaccgc ggattttgtc
     4561 cgttcctctc tctgcgccct cagcgttccc cacactctct cgcttctctt ttcctagacg
     4621 tcttcttttt tcatcttctt cagccctttt tcgccatttt ccatctctgt caatcattac
     4681 ggacgacccc cattaatttt tggatgtgct ctgcgagcaa gagcacacgg aactcattta
     4741 ttttttgttg gaatttcttt ttcattttag ttagttaaaa ggttattcag tgcaaaaatt
     4801 gattttttgc tcttttggta cattagaggg gtttgaaagt tttccataaa aagggtttcc
     4861 ggagtaaaaa atctaaaaat ttttacaaac atttccaaaa attttgggaa catatctttt
     4921 caaaaactca aaaaaaaacg caaacatcat aaatttaatc aattaattgg atgcgttcaa
     4981 atattttaaa aatatttttg aattgcatat ttctttactc ataaaaaaat tattaatgtt
     5041 taagtaaaaa aaatatttca ggattttaaa caaaaactgt aaatcccata aaacagtgtt
     5101 attttttagt tatttttagt aattatttgt gaattaaaaa cacatttttt ttcaaaatat
     5161 tccttgaata aataaaattg aaagtgaatt caaaatattt caccaatttt acaaaatttg
     5221 aaaatttgaa aaaatatttt ctaccaaatt gtgctcaaaa aatattatta agttcccagg
     5281 aaatttatga tgaaaacttg gaatattttt ggtaaaattt tctcaaaaaa ttgaaagttg
     5341 tcaaaaacac actaacacta tacctcgcgg catagaaaaa actggtggcc gaatttttta
     5401 aaccaattta aattaaatat atttttgcac tttgtacaaa aaaatttgaa gttcccagga
     5461 aattcatacg gaaatatttt ccagaacttt tataattttt attttaattc tcaaaattta
     5521 caataaaaac ttggaaacct gaaatttgct taaagaataa ccctatcccc cacggcctag
     5581 aaaatactgg tggccgaatt ttttccgcag ccacaccacc tctaatctcc agagcactct
     5641 ccaaaattcc cataatcccc aaaattttca gctacaacag catatatgaa ggcagagcaa
     5701 caacaacaat cgatcgcacc cggctcggca accttcccgt ctcaggtgag gaagacgcag
     5761 agaaatgaga gagagagcgt gtttatgcac gcagtatact aactcctgat atccaatttc
     5821 atatatcgat atgcttgctt tcttttcttc tacgttgaaa ttgtttgaac ttttctcaaa
     5881 aaacgcggat tttagagaga cttagagttg ctccaaataa atttaatttg caattttgca
     5941 cttactatta gaatctactt ttatagcgct aaaatgaccg aataacacag tagattttaa
     6001 aagaaaattt taaaaattat ttttttgtgc agtcaaattt cgctagtttt tgccactcat
     6061 ttttgaacat ttgtatatgt gcctttcggt ggttcaatca ggggtggcac gtggtgtcag
     6121 agtgttccat ttcggtttga tctacgttga tctacaaaaa atgcgggaga agagacgcag
     6181 agttctcaac tgatttcaca tggttatgaa ccaggggtgt tcggcaaatt tgccgtttgc
     6241 cgagctgggc aaattttgag atttgccgca cacccctttt atgaacgtgc tgacgtcaca
     6301 tctttttgga caaagaattc ccgcattttt tgtatatcaa accgtgatgg gacagcctgg
     6361 caccacgtgg ggttggcggc aaatgatttt ttccgacaaa tcggcaaatt ggcaaattgc
     6421 cggaatttaa agtttctggc aaattgttga tttgccgaaa aaaaatttca aaaaggcaca
     6481 gttttaagtg tttccgtctt ataaaatatt tctctaaaaa tttccggaaa acggaaaatt
     6541 gccggaaata caaagtttcg tcaaatcggc aaacctgcaa tttgccgaac ggcaactgcc
     6601 gccaacacct ggttcccaaa aaagcgtgca ctgtttgaat attaaaaaaa aaattcacac
     6661 aaaaataata aattgaaata ggctaaattc acatctatgt acaaaaattg catgccagca
     6721 ccggtcaatt ttctcaatta ctttttgtct atttttgccg aacataattt gctgtttcct
     6781 tggcagaata cagaaaactc tttttttttt cagatgcccc gacccccacc ggtaaccgaa
     6841 caggctataa ccaccgaggc ggagcttctc gcgaaaaatc aaattacacc aaatgatgtg
     6901 ctagctcttc caggaatcac tcaaggtaag tgaatatttg ctctattgat aaacgcgccg
     6961 tgtactccac gtggacaaac atatcaaatt ttcaggattc ctatgctccc catcggcaaa
     7021 cgtctataac attgagttca ccaaattcca aatccgtgac ctcgacacgg aacacgtgct
     7081 cttcgaaatt gcaaaaccgg aaaatgagac ggaagagaat ctgcaggcgc aagccgaatc
     7141 ggcaagatat gtccgatatc gatttgcgcc gaattttctg aaattaaaga cggtcggcgc
     7201 gacggtcgag ttcaaggtcg gcgatgtgcc gatcacacat tttcgaatga ttgaacggca
     7261 ctttttcaag gatcgtttat tgaaatgctt tgattttgaa tttggattct gtatgccaaa
     7321 ttcgagaaac aactgtgaac atatctatga atttccacaa ctttcacaac aactcagtga
     7381 gtttgaagcg ggaaaaatta tttgagctca ttttgtcaat agagcgaatt ttctaggttt
     7441 ttaaaaataa aataaatatt gacaaaaagc tgccctacag tagttcaaaa tgtgaaaact
     7501 tggaggaaat tcgagttttc tctgagaagt ttttggcggg taattcaaat tttctgagaa
     7561 aaaaattagc cgcgagaatt caaattttct gagaaaatta tttgacggga aattcaaaat
     7621 ttctggaaat gtattgaaca tttgagaaat tttgttaaaa actacattaa accgataaaa
     7681 aagttcgcaa atttttttta agtgctcatg gtgcatcgaa gagaagaatt cttgccaaaa
     7741 ctagcccatg aaattaaaaa ttatcgcgga aaaaaactga ctaattggaa agtaaaatga
     7801 aatttgcaaa aaaaaaatat gtttttttgt tgtaaaatta aaaaaaaaac tgtatatttt
     7861 cagtggacga catgatcaat aatccaaacg agacccgctc cgatagcttc tattttgtgg
     7921 agaataagct cgtaatgcac aataaggccg actactcgta tgatgcataa tttcccgccg
     7981 atggaataaa actaataatt tttctcgcaa tttatagaaa tcatgtgatt ttcctaccgc
     8041 taaatttaaa atttcctaat ttttcactat atttctcaca aaatgcattc cgaaaacact
     8101 agtccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgcaa taaaatatct
     8161 ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagtact aacatacgct
     8221 ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat aggctgtccc cagtgcaagt
     8281 gcaggtgcca gaacatttct ctatcgatag gtaccgagct cttacgcgag atcaaagggg
     8341 gtaagatcaa agggggtaag atcaaagggg gtaagatcaa aggcgcgaga tcaaaggggg
     8401 taagatcaaa gggggtaaga tcaaaggggg taagatcaaa ggggcgcgcc cgcgtgctag
     8461 caaaaatgca taaggttttg ctggcactgt tctttatctt tctggcacca gcatccgcac
     8521 tggcagtctc cgaaccggcc tgcaggatcg attttttgca aattacgagc gttgtagggg
     8581 gcggacgata ggtcctatag gttttgtata tcatcattca ttcattcatt ggtacattca
     8641 tttacccacc ttcctctttc tgagcttctc tggagttctg tgcttccttt ttcccttatc
     8701 tttatactgt aatttttaac tttcaggcat tgattgc
//
