LOCUS       sequence-392393-        6190 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 6190)
  AUTHORS   Coffinier C, Ketpura N, Tran U, Geissert D, De Robertis EM
  TITLE     Mouse Crossveinless-2 is the vertebrate homolog of a Drosophila 
            extracellular regulator of BMP signaling.
  JOURNAL   Mech Dev. 2002 Dec;119 Suppl 1(Suppl 1):S179-84. doi: 
            10.1016/s0925-4773(03)00113-8.
  PUBMED    14516682
REFERENCE   2  (bases 1 to 6190)
  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: "Mech Dev.";
            date: "2002-12"; volume: "119 Suppl 1(Suppl 1):S179-84. doi"; pages:
            " 10.1016/s0925-4773(03)00113-8"
COMMENT     Sequence Label: pCS2 mouse CV2 PC
FEATURES             Location/Qualifiers
     source          1..6190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             2632..2643
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             3152..3187
                     /codon_start=1
                     /product="epitope tag derived from human Protein C (Stearns
                     et al., 1988)"
                     /label=Protein C tag
                     /translation="EDQVDPRLIDGK"
     polyA_signal    3220..3354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3244..3263)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3298..3317
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3467..3487)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3467..3485)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3506..3522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3506..3522)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3519..3541)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3530..3546
                     /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(3554..3560,3561..3578,3579..3584))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3599..3620
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3737..3754)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3908..4496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3988..4007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4667..5458,5459..5527))
                     /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     5290..5309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5528..5632)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5658..6113)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5795..5816)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6007..6026
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat cccatcgatt cgaattggga
     1081 ttccctccag gtgacgatgc tctggttctt cagcgtcagg gctctggctg agcgtccttg
     1141 ccgacgctcg cctgggatta cctgctgcgt cttgctgctg ctcaattgct ccggggtccc
     1201 catgtctctg gcttcctcct tcctgacagg gtctgttgca aaatgtgaaa atgaagggga
     1261 ggtcctccaa attccattta tcacagacaa cccttgcata atgtgtgtgt gtttgaacaa
     1321 agaggtgact tgtaagaggg agaagtgccc tgtgctgtca cgggactgtg cgctggccat
     1381 caagcagaga ggcgcctgct gtgaacgatg caaaggttgt acccatgaag gaagaaccta
     1441 caacagttcc ttcaagtggc aaaccccggc tgagccatgt gtccttcggc agtgccagga
     1501 gggtgtggtg acagagtctg aggtgcgctg tgttgttcat tgtaaaaatc ccgcagagca
     1561 tcaaggagcc tgctgcccta cctgcccagg ctgtgtgttt gaaggtgtgc aataccggga
     1621 aggagaagaa tttcagccag aaggaaacaa atgtattaca tgctcctgcg ttggaggcag
     1681 gacacagtgc gtgagagaag tctgtcccat tctctcttgt ccccagcacc ttagtcacac
     1741 accctcagga cagtgctgcc ccaaatgttt gggtcagaga aaagtattcg accttccctt
     1801 tgggagctgc ctgtttcgca gtgatgttta tgacaatgga gcctcatttg tctatgataa
     1861 ctgcacagta tgcacttgca aggactcaac aatggtttgt aagaagaagt gctcccaccc
     1921 tggtgtctgt aacagtgatg aggatgcctg ttgtgaagac tgtctcttac gggtgccccc
     1981 tgaagacatc aaagtgtgca agttcggtag caagattttc agggatggag agatgtggtc
     2041 ctcagtcaac tgcagcatct gtgcttgtgt gaaggggaag acagagtgtc gaaagaagca
     2101 gtgtgttcct gtcagcagct gcccacaggg taaaattctc aacaggaaag gatgctgtcc
     2161 tatctgcact gaaaaacctg gtgtgtgcac agtattcgga gatccccact acaacacttt
     2221 tgatgggcgg acatttaact ttcaggggac gtgtcagtat gttttgacaa aagactgctc
     2281 ctccccagcc tcacccttcc aggtgctggt gaagaatgat gcacggagga cccgttcctt
     2341 ctcctggacc aagtctgtgg agctgatgct gggcgaaagc acagtcagcc tgcagcagca
     2401 cctcactgtg cgctggaatg gctctcgcat tgcccttccc tgccacacac cccacttcca
     2461 catcgacctg gacggctacc ttttgaaagt gactaccaga gcaggtttgg agatatcgtg
     2521 ggatggagac agttttgtgg aggtcatggc tgcccctcat ctcaagggca agctctgtgg
     2581 tctttgtggc aactacaatg gtcataaacg tgatgacttg attggtggag atgggaactt
     2641 caagtttgac gtggatgact ttgctgaatc gtggagggtg gaatccaacg aattctgcaa
     2701 ccgaccacag agaaaaccag tacctgagct ctgtcaaggc acagtgaagg tgaagcttcg
     2761 agcccatcga gaatgccaga aactcaagtc ctgggagttc cagacctgcc actctacagt
     2821 ggactatacc actttctacc ggtcctgtgt aacagacatg tgtgaatgtc cagtccataa
     2881 aaactgttac tgtgagtcat ttctggcgta tacccgtgcc tgtcagagag agggaatcaa
     2941 agtgcactgg gaacctcagc agagctgtgc agccacccag tgcaaacatg gtgctgtgta
     3001 tgatacctgt ggcccaggat gtgtgaagac atgtgataac tggaatgaga tcggcccatg
     3061 caataagccc tgcattgcag gttgccactg cccagcaaat ctggtccttc ataagggaag
     3121 gtgcatcaag ccagtcctct gtcctcagcg agaagatcag gtagatccac ggttaatcga
     3181 tggtaagtaa tctagaacta tagtgagtcg tattacgtag atccagacat gataagatac
     3241 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     3301 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     3361 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaattc
     3421 gcggccgcgg cgccaatgca ttgggcccgg tacccagctt ttgttccctt tagtgagggt
     3481 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3541 tcacaattcc acacaacata cgagccggga gcataaagtg taaagcctgg ggtgcctaat
     3601 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3661 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3721 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3781 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3841 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3901 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3961 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4021 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4081 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4141 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4201 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4261 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4321 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4381 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4441 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4501 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4561 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4621 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4681 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4741 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4801 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4861 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4921 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4981 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5041 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5101 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5161 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5221 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5281 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5341 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5401 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5461 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5521 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5581 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5641 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     5701 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     5761 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     5821 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     5881 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     5941 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     6001 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     6061 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     6121 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     6181 ccagtcgacc
//
