LOCUS       sequence-356922-        5817 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 5817)
  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 5817)
  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-mWnt8b
FEATURES             Location/Qualifiers
     source          1..5817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        44..423
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        424..627
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     578..598
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     624..648
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        858..876
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     858..875
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     889..908
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     964..980
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     964..980
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2112..2128)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2113..2129)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     regulatory      2147..2156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2160..2294
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2197..2216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2251..2270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2363..2720
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2363..2554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2386..2406)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2571..2706
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2740..2757)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2740..2756)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2749..2771)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2969..3424
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3056..3075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3266..3287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3441..3460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3560..3582
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3620..3638)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3706..3810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3811..3879,3880..4671)
                     /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(4029..4048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4842..5430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5331..5350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5584..5601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5718..5739
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5754..5759,5760..5777,5778..5784)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5792..5808
                     /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     join(5797..5817,1..2)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(5816..5817,1..15)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5816..5817,1..15)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggaaacagct atgacatgat tacgaattaa ttcgagctcg cccgacattg attattgact
       61 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      121 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      181 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      241 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      301 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      361 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      421 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
      481 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
      541 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
      601 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
      661 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggccgg
      721 gaacggtgca ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga
      781 gtctataggc ccaccccctt ggcttcgtta gaacgcggct acaattaata cataacctta
      841 tgtatcatac acatacgatt taggtgacac tatagaataa catccacttt gcctttctct
      901 ccacaggtgt ccactcccag gtccaactgc acctcggttc tatcccaccg cggtggcggc
      961 cgctctagaa ctagtggatc caacatgctt cccatctctc aatgtttgag tcgctcaccc
     1021 tgcctttctc cgaagaccat gtttcttatg aagcccgtgt gcgttcttct agtcacttgt
     1081 gtccttcacc gcagccacgc ctggtcagtg aacaattttc tgatgaccgg tccaaaggct
     1141 tacctggtct actccagcag cgtggccgct ggcgcccaga gtggtattga agaatgtaaa
     1201 taccagtttg cttgggaccg ttggaattgc cccgagagag ctttacagct gtccagccat
     1261 ggtggacttc gaagcgctaa ccgggagaca gcatttgtgc acgccatcag ctctgctggg
     1321 gttatgtaca ccctgactag aaactgcagc ctcggagact ttgacaactg tggctgtgat
     1381 gactcccgaa atggacaact ggggggccaa ggttggctct ggggaggctg cagtgacaac
     1441 gtgggcttcg gagaggcaat ttccaagcag tttgtggatg ccctcgagac aggacaagat
     1501 gcccgggcag ccatgaatct gcacaacaat gaggctggcc gcaaggcggt caagggcacc
     1561 atgaaacgca cgtgtaagtg ccacggtgtg tccggcagct gcaccacgca gacctgctgg
     1621 ttgcaactgc cagagttccg ggaggtaggc gcgcacttga aggagaagta tcatgcggcg
     1681 ctcaaggtgg acctgctgca aggcgcgggc aacagcgcgg cgggccgcgg agccatcgcc
     1741 gacaccttcc gctccatctc cacccgcgag ctggtgcatc tggaggactc cccagactac
     1801 tgcctggaga acaagaccct ggggctgctg ggcaccgagg gccgagagtg tctgcggcgc
     1861 gggcgcgccc tgggtcgctg ggagcgccgc agttgtcgcc ggctgtgcgg ggactgcggg
     1921 ctagcggtgg aggagcgccg cgccgagaca gtgtccagct gcaactgcaa gtttcactgg
     1981 tgctgcgcgg tccgctgcga gcagtgccgc cggcgggtca ccaagtactt ctgcagccgc
     2041 gcagagcggc cgcccagagg cgctgcgcac aaaccgggaa agaactccta agaattcgat
     2101 atcaagctta tcgataccgt cgacctcgag ggggggcccg agcttggccg ccatggccca
     2161 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     2221 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     2281 atcatgtctg gatcgggaat taattcggcg cagcaccatg gcctgaaata acctctgaaa
     2341 gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     2401 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2461 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2521 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     2581 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     2641 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     2701 gaggcctagg cttttgcaaa aagctgttaa cagcttggca ctggccgtcg ttttacaacg
     2761 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     2821 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     2881 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     2941 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3001 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3061 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3121 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3181 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3241 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3301 cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3361 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     3421 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3481 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3541 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3601 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3661 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3721 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3781 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3841 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3901 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3961 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4021 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4081 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4141 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4201 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4261 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4321 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4381 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4441 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4501 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4561 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4621 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4681 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4741 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4801 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4921 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4981 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5281 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5581 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5641 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5701 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5761 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacaca
//
