LOCUS       sequence-243535-        6626 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 6626)
  AUTHORS   Yu H, Ye X, Guo N, Nathans J
  TITLE     Frizzled 2 and frizzled 7 function redundantly in convergent 
            extension and closure of the ventricular septum and palate: evidence
            for a network of interacting genes.
  JOURNAL   Development. 2012 Dec 1;139(23):4383-94. doi: 10.1242/dev.083352. 
            Epub 2012 Oct 24.
  PUBMED    23095888
REFERENCE   2  (bases 1 to 6626)
  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: 
            "10.1242/dev.083352"; journalName: "Development"; date: "2012-12-1- 
            1"; volume: "139"; issue: "23"; pages: "4383-94"
COMMENT     Sequence Label: pRK5-mFzd10
FEATURES             Location/Qualifiers
     source          1..6626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        144..523
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        524..727
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     678..698
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     724..748
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        958..976
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     958..975
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     989..1008
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1064..1080
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1064..1080
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             2872..2901
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3056..3065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3069..3203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3106..3125)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3160..3179
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3272..3629
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3272..3463
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3295..3315)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3480..3615
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3649..3666)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3649..3665)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3658..3680)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3878..4333
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3965..3984)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4175..4196
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4350..4369)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4469..4491
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4529..4547)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4615..4719
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4720..4788,4789..5580)
                     /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(4938..4957)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5751..6339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6240..6259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6493..6510
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
      121 tacgaattaa ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat
      181 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      241 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      301 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      361 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      421 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      481 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      541 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      601 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      661 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      721 cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct
      781 ccatagaaga caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg
      841 gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt
      901 ggcttcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
      961 taggtgacac tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag
     1021 gtccaactgc acctcggttc tatcccaccg cggtggcggc cgctctagaa ctagtggatc
     1081 ccccgggctg caggaattcc caccatgcaa cacccgggcc cgcgcctgtg gctggtgctg
     1141 caggtgatga taggctcgtg cacggccatc agctccatgg acttagggcg ccctggagac
     1201 ggcaagtgcc agccggtgga gattcccatg tgcaaggaca tcggctacaa caccacccgc
     1261 atgcccaacc tgatgggtca cgagaaccag cgcgaggcgg ccatccaact gcacgagttc
     1321 gcgccgctcg tggagtacgg ctgccacagc caccttcgct tcttcctgtg ttcgctgtac
     1381 gcgcccatgt gcaccgagca ggtctccaca cccatccctg cttgccgggt catgtgcgag
     1441 caggcccggc tcaagtgctc gccgatcatg gagcagttca aattcaggtg gccggactcc
     1501 ctggattgca gcaagctccc caacaagaac gaccccaact acctgtgcat ggaggcaccc
     1561 aacaacggct cggatgagcc cagccggggc tctggcatgt ttcctccgct cttcaggccc
     1621 cagaggcccc acagcgcgca ggagcaccca ctaaaggacg ggggtccggg gcgcgcaggt
     1681 tgtgacaacc caggcaagtt ccaccatgtg gagaagagcg aatcttgcgc accgctttgc
     1741 actccggggg tggatgtgta ttggagccgc gacgacaagc gcttcgctgt ggtctggctg
     1801 gccatctggt ccgtgctgtg cttcttctcc agcgccttca ccgtgctcac cttcctcatc
     1861 gacccatcgc gcttcaggta ccccgaacgt cctatcatct tcctctccat gtgctactgc
     1921 gtttattcgg tgggctatat catccgcctc ttcgcgggcg cggagagcat cgcttgtgac
     1981 cgggacagtg ggcagctgta tgttatccag gaaggactgg agagcacggg ctgtacctta
     2041 gtcttcttgg tactttacta cctcggcatg gccagctctt tatggtgggt ggtcctcacc
     2101 ctcacttggt tcctggctgc tggaaagaag tggggccatg aggccattga agccaacagc
     2161 agctactttc acctggcagc ttgggccatc cctgctgtga agactatctt gatcttggtg
     2221 atgcgcaggg tggcagggga tgagctcact ggtgtgtgtt atgtgggcag catggatgtc
     2281 aatgctctga ccggcttcgt gctggtcccg ctggcttgct acctagtcat cggcacttcc
     2341 ttcatcctgt ccggctttgt ggctttattc cacatccgga gggtgatgaa aacgggtggg
     2401 gagaacacgg acaagctgga gaagctcatg gtacgcatag gggtcttctc cctcctctac
     2461 actgtgccgg ccacctgtgt gattgcctgc tatttttatg aacgcctcaa catggactac
     2521 tggaagatgc tggccaccca gcacaagtgt aagatgaaca atcagaccaa gacacctgac
     2581 tgcctgatga ccacctccat ccctgccgtg gaggtcttca tggtcaaagt gtccatgctg
     2641 ctggtggtgg gcatcaccag tggggtgtgg gtctggactt ccaagaccct gcagtcctgg
     2701 caacacgtat gcagccgggg gctaaagaga aaaagccgga ggaaaccagc cagtgtggtc
     2761 accagtgcag ggatctacaa aaaagcccag cacccccaaa aacctcacct tgggaagtat
     2821 gaactgcccg cccagccttc agcctgcgtg tgaaagcttg gcggaggaat ggaacagaag
     2881 ctaatctcag aagaagacct gaacggtgga ggactcgagg aaacaacccc aaataaagga
     2941 agtggaacca cttcaggtac tacccgtctt ctatctgggc acacgtgttt cacgttgaca
     3001 ggtttgcttg ggacgctagt aaccatgggc ttgctgactt aggggcccga gcttggccgc
     3061 catggcccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3121 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3181 atgtatctta tcatgtctgg atcgggaatt aattcggcgc agcaccatgg cctgaaataa
     3241 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg
     3301 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3361 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3421 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3481 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3541 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3601 gcttttttgg aggcctaggc ttttgcaaaa agctgttaac agcttggcac tggccgtcgt
     3661 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     3721 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     3781 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg
     3841 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
     3901 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3961 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4021 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4081 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4141 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4201 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
     4261 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4321 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     4381 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4441 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4501 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     4561 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4621 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4681 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4741 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4801 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4861 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4921 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4981 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5041 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5101 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5161 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5221 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5281 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5341 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5401 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5461 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5521 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5581 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5641 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5701 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5761 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5821 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5881 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5941 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6001 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6061 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6121 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6181 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6241 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6301 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6361 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6421 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6481 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     6541 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     6601 aaagcgggca gtgagcgcaa cgcaat
//
