LOCUS       sequence-242261-        7830 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 7830)
  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 7830)
  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-mFzd2
FEATURES             Location/Qualifiers
     source          1..7830
                     /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"
     regulatory      4260..4269
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4273..4407
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4310..4329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4364..4383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4476..4833
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4476..4667
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4499..4519)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4684..4819
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4853..4870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4853..4869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4862..4884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5082..5537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5169..5188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5379..5400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5554..5573)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5673..5695
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5733..5751)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5819..5923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5924..5992,5993..6784)
                     /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(6142..6161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6955..7543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7444..7463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7697..7714
                     /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 tatcgattga attccaccat gcgggcccgc agcgccctgc
     1081 cccgcagcgc cctgccccgc ctgctgctgc cattgctgct gctgccggcc gccgggccgg
     1141 cccagttcca cggggagaag ggcatctcca tcccggacca cggcttctgc cagcccatct
     1201 ccatcccgct gtgcacggac atcgcctaca accagaccat catgcccaac cttcttggcc
     1261 acacgaacca ggaagacgcg ggcctggagg tgcatcagtt ctacccgctg gtgaaggtgc
     1321 agtgctcgcc cgagctgcgc ttcttcctgt gctccatgta cgcgccggtg tgcacagtgc
     1381 tggagcaggc catcccgccg tgccgctcca tctgcgagcg cgcgcgccaa ggctgcgagg
     1441 cgctcatgaa caagttcggc ttccaatggc ccgagcgcct ccgctgcgag catttcccgc
     1501 gtcacggcgc ggagcagatc tgcgtgggcc agaaccactc ggaggacgga gctcctgcgc
     1561 tactcaccac cgcgccacct tctgggctgc agcccggcgc gggtggcacc ccgggcggcc
     1621 ctggcggtgg tggcgcgcca ccgcgttacg ccactctgga gcaccctttc cactgtcccc
     1681 gcgtcctcaa ggtgccgtcc tatctcagct ataagtttct gggtgagcgc gattgtgccg
     1741 cgccctgcga gcctgcacgg cccgacggct ctatgttctt ctcgcaagag gagactcgtt
     1801 ttgcccgtct ctggatcctc acatggtcgg tgctgtgctg cgcttccacc ttcttcacgg
     1861 tcaccaccta tttagtggac atgcagcgat ttcgctaccc agagcggccc atcatctttc
     1921 tgtccggctg ctacaccatg gtgtcagtgg cctacattgc gggcttcgtt ctccaggagc
     1981 gcgtggtatg caatgagcgc ttctcagagg acggttatcg cacggtggtg cagggcacta
     2041 agaaagaagg ctgcactata ctcttcatga tgctctactt cttcagcatg gccagctcca
     2101 tctggtgggt gattctgtcc ctcacctggt tcctggcagc cggaatgaag tggggccacg
     2161 aggccatcga ggccaattcg cagtacttcc acctggccgc ctgggccgtg ccggccgtca
     2221 aaaccatcac catcttggcc atgggccaga tcgacggcga cctgctgagc ggcgtgtgct
     2281 tcgtgggcct caatagcctg gacccgctgc ggggcttcgt gctggcgccg ctcttcgtat
     2341 acctgttcat cggtacatcc ttcctgctgg ccggcttcgt gtcactcttc cgcatccgca
     2401 ccatcatgaa gcacgacggc accaagacgg agaagctgga gaggctcatg gtgcgcattg
     2461 gcgtcttctc ggtgctctac acggtaccgg ccaccatcgt catcgcctgc tacttctatg
     2521 agcaggcctt ccgcgagcac tgggagcgct cctgggtaag ccagcactgc aagagcctag
     2581 ccatcccctg cccggcccac tacacgcccc gcatgtcgcc cgacttcaca gtctacatga
     2641 tcaaatacct catgacgctc atcgtgggca tcacgtcggg cttctggatc tggtccggca
     2701 agacactgca ctcgtggagg aagttctaca cgcgtctcac caacagccgg catggcgaga
     2761 ccactgtgtg aagcggtctc gcctgcctgc cgggcgttcc cctctcccag gtccggactg
     2821 caccgtgccc tccttcactc aggagggggg agggtgcacc ctacggactc ctattttatt
     2881 tttttaaata aagaacggta aaaaaaaaac attttatttt taggtcgctt tttaaaaaga
     2941 actctgccca acacccccct accaggtttg taattaaaac tgtaaatagt ctttgtaaat
     3001 ttaattatat attttctatt taaaagaaaa aagaaaaaaa aatcgaagac accagggctg
     3061 aagagtggtg tgggggtggg ggaggaggtc tcgacttttc taagtgccct tttaaaagct
     3121 gccctgtgct ctggttcgaa ttttttttag tgatttggca gggctgggcc tgctgggaga
     3181 ggcacctagc ctgctcgcta tttttgttgg cttaaaggag ttgggagtta gttccactca
     3241 acttctataa aagctaaatt tgcaagcctc ctcaccgacg aggggcctgt ggaagctgtt
     3301 ggatatattt tttaaacaag acttggattc gtttttgttt tatttccctt tctcctcgct
     3361 ccacaccccc catccccctc tttctccctc ctcatttgtt ctgtctcccc cgctcgctca
     3421 gtatctggaa acctcccaat ccagatgtag acgtggaaaa aaacatagcg cttttaggga
     3481 gagggcggag atgtggggag gaatggcctc caccccccac ccccagccca ctccccgtgg
     3541 gcacggagat ccttctgtgg tttcagctct ggcccacgcg cagagaatgg gcaacaaaac
     3601 gctttgtgta gtgtgtcttg acatcccccc ccccagtgca cacaccactt taagaatgac
     3661 ctcctttccc tgggttttgg gggtaccgct agcctctgtg ttggcaaacc ctgggatctt
     3721 tggcgcttcc gttcccttct ctcccggtag gctgctcttc cgcggtggga tccaggttag
     3781 ctgtttgtag gattgggttg tgtgttatgt gtgggttttt gttcgggttg tttttctttt
     3841 ccaaaagcag taaaaatagg ttgggttgga gggagggaat ggatgggctg gtgggatttt
     3901 tagttttctt ttgacaaaag actggccttn aaaaaaaaaa aaaaacaaaa aacaaaaaac
     3961 aaaagcaaaa agttgttcag ggaaacaaca aaaactaatg ggtaagtttg ctttagaagg
     4021 tctctctggg caggctcttt gtggcggtgg gggtggggtt gagtgtttac gtgacattct
     4081 atatcataag atattgataa gaatgtttct aacatgtttc ttcccccccc ccacccccct
     4141 gcacacaaat aaaagtaagg attttgctaa gttaaaggcc tgagagatgt tctaaaaagt
     4201 atctttgtta gtgtgagaat cccaatttcc agcaggcatg ggttccctcg agaagcttgg
     4261 ccgccatggc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4321 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4381 atcaatgtat cttatcatgt ctggatcggg aattaattcg gcgcagcacc atggcctgaa
     4441 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     4501 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4561 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     4621 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     4681 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4741 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4801 ggaggctttt ttggaggcct aggcttttgc aaaaagctgt taacagcttg gcactggccg
     4861 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     4921 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     4981 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     5041 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     5101 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5161 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5221 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5281 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5341 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5401 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     5461 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     5521 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     5581 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5641 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5701 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     5761 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     5821 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5881 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     5941 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     6001 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     6061 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     6121 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     6181 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     6241 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     6301 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     6361 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     6421 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     6481 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     6541 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     6601 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     6661 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     6721 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     6781 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     6841 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     6901 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     6961 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     7021 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     7081 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     7141 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     7201 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     7261 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     7321 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     7381 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     7441 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     7501 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     7561 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     7621 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     7681 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     7741 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     7801 ctggaaagcg ggcagtgagc gcaacgcaat
//
