LOCUS       sequence-264810-        4789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gata2a endothelial enhancer (x6) and basal promoter driving 
            nuclear-localized sfGFP in pTol1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4789)
  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 4789)
  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: pTone-gata2aECE-nsGFP
FEATURES             Location/Qualifiers
     source          1..4789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    53..73
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    412..436
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      439..448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             451..471
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             499..528
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             538..567
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             577..606
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             616..645
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             655..684
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             718..747
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1411..1458
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     protein_bind    complement(1514..1538)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1564..1584)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    1606..1740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1630..1649)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1684..1703
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1941..1957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1993..2009)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2006..2028)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2017..2033
                     /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(2041..2047,2048..2065,2066..2071))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2086..2107
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2224..2241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2395..2983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2475..2494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3154..3945,3946..4014))
                     /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     3777..3796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4015..4119)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4187..4205
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4243..4265)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4365..4384
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4578..4600
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4592..4609
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4593..4609
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cattgggatg gaaccctcga ggggcccgct agcgatatcc ctgcagggct atcaactttg
       61 tatagaaaag ttgagttgtt tcctatccgg atatctgccc gggcccgata aagttgtttc
      121 ctatccggat atctgcccgg gcccgataac acgtcacgca cgtgcagtac tagttgtttc
      181 ctatccggat atctgcccgg gcccgataaa gttgtttcct atccggatat ctgcccgggc
      241 ccgataacac gtcacgcacg tgcagtacta gttgtttcct atccggatat ctgcccgggc
      301 ccgataaagt tgtttcctat ccggatatct gcccgggccc gataagcttc tagagggtat
      361 ataatggatc atcgcgtctc agcctcactt tgagctcctc cacacgctcg agcaagtttg
      421 tacaaaaaag caggctccgc caccatggct cctaagaaga agaggaaggt ggcaggatcc
      481 catcgattta aagctatgga gcaaaagctc atttctgaag aggacttgaa tgaaatggag
      541 caaaagctca tttctgaaga ggacttgaat gaaatggagc aaaagctcat ttctgaagag
      601 gacttgaatg aaatggagca aaagctcatt tctgaagagg acttgaatga aatggagcaa
      661 aagctcattt ctgaagagga cttgaatgaa atggagagct tgggcgacct caccatggag
      721 caaaagctca tttctgaaga ggacttgaat ttagagatct cgaattccat gtccaaagga
      781 gaagagctgt tcactggagt ggtaccaata cttgtggagt tggacggaga tgtgaacgga
      841 cacaaatttt cagtccgcgg ggagggggaa ggggatgcta ctattggcaa gctgacgctc
      901 aaattcatct gtaccaccgg aaaactccct gtaccctggc ccacactggt gacaactctg
      961 acttacggcg tgcaatgttt tagccgatac ccagaccaca tgaagaggca cgactttttc
     1021 aaaagcgcaa tgcctgaagg atacgtacag gaaaggacca tttcttttaa agacgacggg
     1081 aagtacaaaa cccgggcagt ggtgaagttt gagggcgata ccctcgtcaa taggatcgaa
     1141 ttgaagggaa ctgacttcaa agaagatggc aacatcctgg gtcacaagct tgagtataac
     1201 tttaactccc acaacgtgta tattacagcc gacaaacaga agaatggaat taaggctaac
     1261 ttcactgtca gacacaatgt cgaagatggc tccgtgcagc tcgccgatca ctatcaacag
     1321 aatactccta tcggggacgg cccagtcctg ctgcccgaca accactacct gagtacccag
     1381 actgttctga gcaaagatcc gaacgagaag cgcgaccaca tggtgctgca tgagtatgtc
     1441 aacgctgcgg gaattaccct cggcatggac gagctgtaca agtctagagc tccaaagaag
     1501 cgtaaggtat aacacccagc tttcttgtac aaagtggggc gcgcctacgt aactagtctc
     1561 gagcaacttt attatacata gttgataagg cgcgccaggc ctgtagatcc agacatgata
     1621 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1681 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1741 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1801 taattaaaac atttcagttg acgaagacaa acaaagttct gttgtgacta tggggggggg
     1861 gggcgcctgg ggatggtctc gcccggggag taattcaggg tagaaccgcc actggctaaa
     1921 ggaagcttgg cgtaatcatg gtcatagctg tttcctgtat cccaatggcg cgccgagctt
     1981 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2041 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2101 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2161 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2221 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2281 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2341 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2401 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2461 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2521 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2581 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2641 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2701 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2761 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2821 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2881 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2941 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3001 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3061 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3121 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3181 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3241 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3301 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3361 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3421 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3481 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3541 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3601 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3661 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3721 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3781 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3841 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3901 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3961 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4021 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4081 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4141 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4201 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4261 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4321 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4381 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4441 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4501 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4561 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcccttt
     4621 agccagtagc ggttctaggc acgggccgtc cgggcggtgg cctggggcgg aaaactgaag
     4681 gggggcggca ccggcggctc agccctttgt aatatattaa tatgcaccac tattggttta
     4741 cttatgtcac agtttgtaag tttgtaacag cctgaacctg gccgcgcgc
//
