LOCUS       sequence-178755-        7860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  contains tmem88a enhancer sequence, and tmem88a specific gene 
            promoter driving expression of eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7860)
  AUTHORS   Quillien A, Abdalla M, Yu J, Ou J, Zhu LJ, Lawson ND
  TITLE     Robust Identification of Developmentally Active Endothelial 
            Enhancers in Zebrafish Using FANS-Assisted ATAC-Seq.
  JOURNAL   Cell Rep. 2017 Jul 18;20(3):709-720. doi: 
            10.1016/j.celrep.2017.06.070.
  PUBMED    28723572
REFERENCE   2  (bases 1 to 7860)
  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.celrep.2017.06"; journalName: "Cell Rep"; date: 
            "2017-07-18- 18"; volume: "20"; issue: "3"; pages: "709-720"
COMMENT     Sequence Label: pT2-cryR;tmem88aCE1-P1Egfp
FEATURES             Location/Qualifiers
     source          1..7860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     637..653
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    673..693
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    1213..1237
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1876..1885
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1882..1884,1885..1887,1888..2598)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1882..1884,1885..1887,1888..2598)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1927..1948)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2188..2207)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2535..2556
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2599..2664
                     /label=MCS
                     /note="multiple cloning site"
     protein_bind    complement(2685..2709)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2735..2755)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     primer_bind     complement(2763..2779)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2795..2929
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    join(3368..3555,3556..3567)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(3601..3617)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3601..3617)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3654..3674)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3654..3672)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3693..3709)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3706..3728)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3717..3733
                     /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(3741..3747,3748..3765,3766..3771))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3786..3807
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3924..3941)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4095..4683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4175..4194)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4854..5645,5646..5714))
                     /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     5477..5496
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5715..5819)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5982..6003)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     misc_feature    join(6352..6363,6364..6526)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     polyA_signal    6895..7016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(7139..7849)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEDDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGHHSTGGMDELYK"
     primer_bind     complement(7219..7242)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(7417..7436)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7686..7704
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7846..7855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gtaatgtcag acctggtaac tccttactgt aacttattgc acctgatgaa caaaagatgt
       61 gaactgaacg caagtaggag gctaatagag gctagtacag tacaaagatc cccaaatgat
      121 gtgtgccggc cttagtggag gcagacccct ggttatatac ctgtgctgca gaaagagggg
      181 cttacaaatg gatctctctg gaatggcatt atctcatggt gggaacagtg ttgtcagcag
      241 aatcagcggg gtggactctg acctgagaat gacagtctag cactgaatgg ctcagacaat
      301 actcttgtgg ccaactatca ccggaaaata agtgcctgcc aaaacaaacc agaccagatg
      361 aagctacatg gctgtagaac gtcagtattt aatgtgttgt ggccacttgg ttcaccagat
      421 ttaagagcta atttttttaa atatcacaga taatcaatcg cttttttttt tcttgactga
      481 aaagcttaga ctggaaaatt ctttgaaaaa gtttgcacta cagaaaataa atacataaca
      541 tcctgcactg aatgcacact attaatagaa aagagagcat tattgtagga aatgtattgt
      601 atatgtacat tctgtgcatt tcacaatttt ctcgagcagg aaacagctat gaccatgatt
      661 acgccaagct atcaactttg tatagaaaag ttgatttttg agttattatt cctgatataa
      721 tgattactat aaaacaagac taaaagcttc tgagtgttta cgtgtaagtt atttgtggat
      781 tgtgaagtgt attagaggaa ggttacagat ggtcagtctc ctgtctgtct gtgggctgga
      841 agtgaggctc gtagaggtca tgggacagga ctaatgtttt ggtgacacct cggctctctc
      901 tcaggacttt actgcggaaa tcctggctcc aaggccagga aggcaggttt tcctctttta
      961 tgactctcag gatatatctc gtttttcacc gtccctctgc acttaacgct tatgttccac
     1021 cagcagccct ttatctgtgg ctgctgataa cccacagctg tgagtaaata aataaacaac
     1081 tgtcaatagc ttatttactt acagataaag tcgccatcct gtttttcact cttatgaact
     1141 cgatatgttg ttttgttgac tctatcacag tcgttcacaa ctggcaggcc gtgacctgaa
     1201 aaacattcaa aggcaagttt gtacaaaaaa gcaggctcgc taaattaaca acttctttcc
     1261 caacacctaa aatatgcaca ttttcccatg aaatcttggc taaactacac attttgtcag
     1321 tttgtgtgtg tatctgtaac actttacagt aaagttgaac attgtaaaaa gcgattaggt
     1381 tctgtacttc aaaattcaga ttacttcatt ttactttaag aaaagtgagt aataatgtaa
     1441 gtatatctga aatgtattag caatttctga gcagaaataa aacatactgt ggagtgtgta
     1501 actctaaagt cagtgcaagt catagatttt cctactgcat gtgcaaaaaa ataaaaaagt
     1561 ctacaactaa acccccagca aacccaacac cccacatttc tgaagggctg atccagatcg
     1621 ggaggccagg aggagaagtg ggagtggaga gatacgagtg agttactgag gcaggacatc
     1681 ttcgtcaaag gaagaaaaga gtagagcagg aaagaaggaa gagaacgagc tgacatagaa
     1741 gactgacgct cattgagctc acagagcctg aggagatact acaggtggac ctgaaaactg
     1801 gataaaaatc aagtcaaact aactgtagag taaagctatc atctgccagt ctgaggaatc
     1861 tgcccatatt tcatcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1921 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1981 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     2041 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2101 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2161 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2221 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2281 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2341 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2401 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2461 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2521 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2581 atggacgagc tgtacaagtc cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
     2641 gacggtaccg cgggcccggg atccaccgga tctagataac tgatacccag ctttcttgta
     2701 caaagtgggg cgcgcctacg taactagtct cgagcaactt tattatacat agttgataat
     2761 tcactggccg tcgttttacg gtaccatcga tgatgatcca gacatgataa gatacattga
     2821 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2881 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2941 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     3001 aaacctctac aaatgtggta tggctgatta tgatcctcta gatcagatct gcgaagatac
     3061 ggccacgggt gctcttgatc ctgtggctga ttttggactg tgctgctcgc agctgctgat
     3121 gaatcacata cttcctccat tttcttccac tgattgactg ttataatttc cctaatttcc
     3181 aggtcaaggt gctgtgcatt gtggtaatag atgtgacatg acgtcacttc caaaggacca
     3241 atgaacatgt ctgaccaatt tcatataatg tgaaaacgat tttcataggc agaataaata
     3301 acatttaaat taaactgggc atcagcgcaa ttcaattggt ttggtaatag caagggaaaa
     3361 tagaatgaag tgatctccaa aaaataagta ctttttgact gtaaataaaa ttgtaaggag
     3421 taaaaagtac ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt attgattttt
     3481 aattgtactc aagtaaagta aaaatcccca aaaataatac ttaagtacag taatcaagta
     3541 aaattactca agtactttac acctctggtt cttgaccccc taccttcagc aagcccagca
     3601 gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg ttccctttag
     3661 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3721 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3781 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3841 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3901 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3961 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4021 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4501 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4621 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4741 taaggcattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4801 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4861 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4921 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4981 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5041 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5101 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5161 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5221 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5281 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5341 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5401 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5461 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5521 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5581 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5641 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5701 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5761 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5821 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     5881 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     5941 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6001 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6061 gcccacgctg gctaagaact catcagcctc cccggtccat ctacccacgt accaatgcac
     6121 caattggcca caatgacggc tactacatgg tgccattcct tcctctttat aggaatggag
     6181 actacctcct gtccaacaat gctcttggat acgagtacgc ctacctgttg gacccaggtc
     6241 attgcacaac accagaaatg ccctctgatc tgcaaaagac gtgaatatct gttcagacac
     6301 ccatatccac tctgttccac acaggtcaga ggtttgtcca ggagttcttg acagaggtgt
     6361 aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa aattttactc
     6421 aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa aagtactcca
     6481 ttaaaattgt acttgagtat taaggaagta aaagtaaaag caagaaagaa aactagagat
     6541 tcttgtttaa gcttttaatc tcaaaaaaca ttaaatgaaa tgcatacaag gttttatcct
     6601 gctttagaac tgtttgtatt taattatcaa actataagac agacaatcta atgccagtac
     6661 acgctactca aagttgtaaa acctcagatt taacttcagt agaagctgat tctcaaaatt
     6721 gttagtgtca agcctagctc ttttggggct gaaaagcaat cctgcagtgc tgaaaagcct
     6781 ctcacaggca gccgatgcgg gaagaggtgt attagtcttg atagagaggc tgcaaatagc
     6841 aggaaacgtg agcagagact ccctggtgtc tgaaacacag gcccatttac gccttaagat
     6901 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     6961 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     7021 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     7081 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tctagagtcg cggccgcttt
     7141 acttgtacag ctcgtccatg ccgccggtgg agtggtggcc ctcggcgcgc tcgtactgtt
     7201 ccacgatggt gtagtcctcg ttgtgggagg tgatgtccag cttgatgttg acgttgtagg
     7261 cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctcg gcgtcgtagt
     7321 ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg
     7381 ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt
     7441 cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg
     7501 aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact
     7561 tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt
     7621 aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc
     7681 cgcccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct
     7741 cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca
     7801 tgaactcctt gatgatggcc atgttgtcat cctcgccctt gctcaccatg gtggcgaccg
//
