LOCUS       sequence-264809-        7428 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full-length of foxc1a in frame with viral 2a peptide and nuclear 
            localized tdTomato in pCS expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7428)
  AUTHORS   Shin M, Nozaki T, Idrizi F, Isogai S, Ogasawara K, Ishida K, Yuge S,
            Roscoe B, Wolfe SA, Fukuhara S, Mochizuki N, Deguchi T, Lawson ND
  TITLE     Valves Are a Conserved Feature of the Zebrafish Lymphatic System.
  JOURNAL   Dev Cell. 2019 Nov 4;51(3):374-386.e5. doi: 
            10.1016/j.devcel.2019.08.019. Epub 2019 Sep 26.
  PUBMED    31564611
REFERENCE   2  (bases 1 to 7428)
  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: 
            "10.1016/j.devcel.2019.08.019"; journalName: "Dev Cell"; date: 
            "2019-11-4- 4"; volume: "51"; issue: "3"; pages: "374-386.e5"
COMMENT     Sequence Label: pCS-foxc1a-2a-nlstdTomato
FEATURES             Location/Qualifiers
     source          1..7428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        39..1016
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1051..1069
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1051..1068
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1124..1148
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2586..2610)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2621..2677
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2705..2734
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2744..2773
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2783..2812
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2822..2851
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2861..2890
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2924..2953
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2978..2998
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2999..4426
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGSHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(4436..4456)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    4496..4630
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4520..4539)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4574..4593
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4743..4763)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4743..4761)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4782..4798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4782..4798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4795..4817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4806..4822
                     /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(4830..4836,4837..4854,4855..4860))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4875..4896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5013..5030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5184..5772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5264..5283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5943..6734,6735..6803))
                     /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     6566..6585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6804..6908)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6934..7389)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7071..7092)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7283..7302
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat
       61 ggactcatgc caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat
      121 ggactcgtgc caattcaata tggtggatct ggaccccagc caattcaata tggcggactt
      181 ggcaccatgc caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact
      241 tggcacggtg ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat
      301 tggcatggtg ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa
      361 tatccaatat ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc
      421 ctatgccata tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat
      481 gggcggtccc atataccata tatggggctt cctaataccg cccatagcca ctcccccatt
      541 gacgtcaatg gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga
      601 cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc
      661 ccattgacgt caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc
      721 atatccgttc tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac
      781 atcaatatct attaatagta acttggcaag tacattacta ttggaagtac gccagggtac
      841 attggcagta ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac
      901 atccccattg acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa
      961 tgggcggtag gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaaccgcc
     1021 attctgcctg gggacgtcgg agcaagcttg atttaggtga cactatagaa tacaagctac
     1081 ttgttctttt tgcaggatcc catcgattcg aattcaagga tcaacaagtt tgtacaaaaa
     1141 agcaggctcc gccaccatgc aggcgcgcta ttccgtctcc agtcccaact cgctgggagt
     1201 tgtgccgtac atcagcagcg accaaagtta ttaccgagcc gccgccggag gggggtacac
     1261 cggcatgcca gcccccatga ccatgtactc tcacgcggcc cacgaccagt accccgccag
     1321 catggcgagg gcatatgggc catacactcc acagccgcag cctaaggata tggtgaaacc
     1381 cccgtacagc tacattgcgc tcatcacaat ggcaatccag aactccccgg acaaaaaggt
     1441 gacccttaat ggtatttatc agttcatcat ggagcgcttt cccttttaca gagacaacaa
     1501 gcaaggctgg cagaacagca tccgacataa tttatccctg aacgagtgct ttgtcaaagt
     1561 tccacgcgat gataagaaac ccggaaaagg cagctactgg accctcgacc cggactcgta
     1621 taatatgttt gagaacggca gctttttgag gcgacgacgg cgatttaaaa agaaggacgc
     1681 catgaaggac aaggaggacc gaggtgttaa agaggcgccg tcccgccagg cgcagccgca
     1741 ggcccgcgag caggagcagt cggtgcctgg ctctcagccc gtgcgcattc aggacattaa
     1801 aaccgagaac ggcacgtcca ctcctccgca ggctgtttcg cccacgctaa gcaccgtccc
     1861 gaagatcgag agccccgaca gcagcagcag catgtcgagc ggcagcccgc acagcatccc
     1921 ttccacccgc tccctcagcc tggatagcgc tggagagcag caccaccagg cccccgcgca
     1981 gggcttcagc gtggacaaca tcatgacctc cctccgcgga tcgccgcaca gctccggaga
     2041 gctcaccccg tctctcgtgg ctccctcgcg gacaggtata acacccacgt tgtccctgaa
     2101 ttattctccc aatcagtcgt ccgtgtacag ctcgccctgc agccagaact ctatctcgac
     2161 tacctccaac gccacctatc actgcaacat gcaggccatg agcttgtacg cgggcggaga
     2221 tcgctcgggc cacttgggga ccaccgctac caccgtggac gaaactctgc ctgattactc
     2281 gatcacgacc accacgtcgt cgttgagcca cgggaatctg agctcggccc aggagggcca
     2341 ccatccgcac caagggcgcc tcgcctcatg gtacctgaac caggccggag acattggcca
     2401 tctaggcgcg acgtacccag cgcagcagca gaacttccat tcggttcgag agatgttcga
     2461 atcgcagaga atcggtctga ataattcgcc ggtaaacggg aataacagct gtcaaatgtc
     2521 gtttcccccg agccagccca tttaccgcac gtcgggggct tttgtgtatg actgcagcaa
     2581 atttcaccca gctttcttgt acaaagtggg aggatccgga gccacgaact tctctctgtt
     2641 aaagcaagca ggagacgtgg aagaaaaccc cggtcctgca ggatcccatc gatttaaagc
     2701 tatggagcaa aagctcattt ctgaagagga cttgaatgaa atggagcaaa agctcatttc
     2761 tgaagaggac ttgaatgaaa tggagcaaaa gctcatttct gaagaggact tgaatgaaat
     2821 ggagcaaaag ctcatttctg aagaggactt gaatgaaatg gagcaaaagc tcatttctga
     2881 agaggacttg aatgaaatgg agagcttggg cgacctcacc atggagcaaa agctcatttc
     2941 tgaagaggac ttgaatttag agatctcgaa ttccatgccc aagaagaaga ggaaggtgat
     3001 ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc gcatggaggg
     3061 ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
     3121 cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg cctgggacat
     3181 cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg ccgacatccc
     3241 cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
     3301 ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca cgctgatcta
     3361 caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc agaagaagac
     3421 catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc tgaagggcga
     3481 gatccaccag gccctgaagc tgaaggacgg cagccactac ctggtggagt tcaagaccat
     3541 ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca ccaagctgga
     3601 catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct ccgagggccg
     3661 ccaccacctg ttcctggggc atggcaccgg cagcaccggc agcggcagct ccggcaccgc
     3721 ctcctccgag gacaacaaca tggccgtcat caaagagttc atgcgcttca aggtgcgcat
     3781 ggagggctcc atgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     3841 cgagggcacc cagaccgcca agctgaaggt gaccaagggc ggccccctgc ccttcgcctg
     3901 ggacatcctg tccccccagt tcatgtacgg ctccaaggcg tacgtgaagc accccgccga
     3961 catccccgat tacaagaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     4021 cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc tccctgcagg acggcacgct
     4081 gatctacaag gtgaagatgc gcggcaccaa cttccccccc gacggccccg taatgcagaa
     4141 gaagaccatg ggctgggagg cctccaccga gcgcctgtac ccccgcgacg gcgtgctgaa
     4201 gggcgagatc caccaggccc tgaagctgaa ggacggcggc cactacctgg tggagttcaa
     4261 gaccatctac atggccaaga agcccgtgca actgcccggc tactactacg tggacaccaa
     4321 gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg agcgctccga
     4381 gggccgccac cacctgttcc tgtacggcat ggacgagctg tacaagggat cctaacaact
     4441 ttattataca tagttgcact cgagcctcta gaactatagt gagtcgtatt acgtagatcc
     4501 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4561 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4621 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4681 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
     4741 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
     4801 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
     4861 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4921 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4981 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5041 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5101 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5161 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5221 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5281 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5341 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5401 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5461 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5521 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5581 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5641 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5701 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5761 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5821 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5881 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5941 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6001 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6061 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6121 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6181 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6241 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6301 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6361 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6421 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6481 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6541 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6601 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6661 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6721 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6781 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6841 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6901 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     6961 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     7021 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     7081 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     7141 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     7201 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     7261 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     7321 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     7381 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggc
//
