LOCUS       sequence-263554-        4407 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gata2a endothelial enhancer (x6) and basal promoter driving kaede in
            pTol1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4407)
  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 4407)
  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-Kaede
FEATURES             Location/Qualifiers
     source          1..4407
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     51..73
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     65..82
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     66..82
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    315..335
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    674..698
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             716..1393
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)"
                     /label=Kaede
                     /translation="MSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVVK
                     EGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIAT
                     NDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALLL
                     KGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLPD
                     NVK"
     protein_bind    complement(1394..1418)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1444..1464)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     polyA_signal    1486..1620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1510..1529)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1564..1583
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1821..1837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1873..1889)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1886..1908)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1897..1913
                     /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(1921..1927,1928..1945,1946..1951))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1966..1987
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2104..2121)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2275..2863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2355..2374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3034..3825,3826..3894))
                     /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     3657..3676
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3895..3999)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4067..4085
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4123..4145)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4245..4264
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
       61 acgttgtaaa acgacggcca gtgaattccc tttagccagt agcggttcta ggcacgggcc
      121 gtccgggcgg tggcctgggg cggaaaactg aaggggggcg gcaccggcgg ctcagccctt
      181 tgtaatatat taatatgcac cactattggt ttacttatgt cacagtttgt aagtttgtaa
      241 cagcctgaac ctggccgcgc gccattggga tggaaccctc gaggggcccg ctagcgatat
      301 ccctgcaggg ctatcaactt tgtatagaaa agttgagttg tttcctatcc ggatatctgc
      361 ccgggcccga taaagttgtt tcctatccgg atatctgccc gggcccgata acacgtcacg
      421 cacgtgcagt actagttgtt tcctatccgg atatctgccc gggcccgata aagttgtttc
      481 ctatccggat atctgcccgg gcccgataac acgtcacgca cgtgcagtac tagttgtttc
      541 ctatccggat atctgcccgg gcccgataaa gttgtttcct atccggatat ctgcccgggc
      601 ccgataagct tctagagggt atataatgga tcatcgcgtc tcagcctcac tttgagctcc
      661 tccacacgct cgagcaagtt tgtacaaaaa agcaggctcg ctaccggtcg ccaccatgag
      721 tctgattaaa ccagaaatga agatcaagct gcttatggaa ggcaatgtaa acgggcacca
      781 gtttgttatt gagggagatg gaaaaggcca tccttttgag ggaaaacaga gtatggacct
      841 tgtagtcaaa gaaggcgcac ctctcccttt tgcctacgat atcttgacaa cagcattcca
      901 ttatggtaac agggtttttg ctaaataccc agaccatata ccagactact tcaagcagtc
      961 gtttcccaaa gggttttctt gggagcgaag cctgatgttc gaggacgggg gcgtttgcat
     1021 cgctacaaat gacataacac tgaaaggaga cacttttttt aacaaagttc gatttgatgg
     1081 cgtaaacttt cccccaaatg gtcctgttat gcagaagaag actctgaaat gggaggcatc
     1141 cactgagaaa atgtatttgc gtgatggagt gttgacgggc gatattacca tggctctgct
     1201 gcttaaagga gatgtccatt accgatgtga cttcagaact acttacaaat ctaggcagga
     1261 gggtgtcaag ttgccaggat atcactttgt cgatcactgc atcagcatat tgaggcatga
     1321 caaagactac aacgaggtta agctgtatga gcatgctgtt gcccattctg gattgccgga
     1381 caacgtcaag taaacccagc tttcttgtac aaagtggggc gcgcctacgt aactagtctc
     1441 gagcaacttt attatacata gttgataagg cgcgccaggc ctgtagatcc agacatgata
     1501 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     1561 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1621 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1681 taattaaaac atttcagttg acgaagacaa acaaagttct gttgtgacta tggggggggg
     1741 gggcgcctgg ggatggtctc gcccggggag taattcaggg tagaaccgcc actggctaaa
     1801 ggaagcttgg cgtaatcatg gtcatagctg tttcctgtat cccaatggcg cgccgagctt
     1861 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     1921 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     1981 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2041 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2101 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2161 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2701 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2941 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3001 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3061 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3121 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3181 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3241 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3301 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3361 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3421 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3481 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3541 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     3601 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     3661 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3721 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3781 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3841 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3901 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3961 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4021 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     4081 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     4141 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     4201 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     4261 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     4321 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4381 ggcctcttcg ctattacgcc agctggc
//
