LOCUS       sequence-243958-        7205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transgene to report estrogen receptor activity in zebrafish.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7205)
  AUTHORS   Gorelick DA, Halpern ME
  TITLE     Visualization of estrogen receptor transcriptional activation in 
            zebrafish.
  JOURNAL   Endocrinology. 2011 Jul;152(7):2690-703. doi: 10.1210/en.2010-1257. 
            Epub 2011 May 3.
  PUBMED    21540282
REFERENCE   2  (bases 1 to 7205)
  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: 
            "10.1210/en.2010-1257"; journalName: "Endocrinology"; date: 
            "2011-07"; volume: "152"; issue: "7"; pages: "2690-703"
COMMENT     Sequence Label: pT2-5xERE:GFP
FEATURES             Location/Qualifiers
     source          1..7205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(15..26)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     regulatory      87..96
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(93..95,96..98,99..812)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(93..95,96..98,99..809)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(138..159)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(399..418)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     746..767
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    857..991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(881..900)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     935..954
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1430..1617,1618..1629)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1663..1679)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1663..1679)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(1716..1736)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1716..1734)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1755..1771)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1755..1771)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1768..1790)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1779..1795
                     /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(1803..1809,1810..1827,1828..1833))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1848..1869
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1986..2003)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2157..2745)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2237..2256)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2916..3707,3708..3776))
                     /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     3539..3558
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3777..3881)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3907..4362)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4044..4065)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4256..4275
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4489..4511
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4503..4520
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4504..4520
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4530..4549
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4530..4548
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(6395..6406,6407..6569)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 cacccagctc tgctctgcag ctcccaccag tgtctacccc tggacccctt gccgggcttt
       61 ccccaaactt cgaggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
      121 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      181 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      241 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      301 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      361 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      421 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      481 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      541 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
      601 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      661 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
      721 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      781 tcactctcgg catggacgag ctgtacaagt aaagcggccg ccaccgcggt ggagctcgaa
      841 ttaattcatc gatgatgatc cagacatgat aagatacatt gatgagtttg gacaaaccac
      901 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      961 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     1021 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     1081 tatggctgat tatgatcctc tagatcagat ctgcgaagat acggccacgg gtgctcttga
     1141 tcctgtggct gattttggac tgtgctgctc gcagctgctg atgaatcaca tacttcctcc
     1201 attttcttcc actgattgac tgttataatt tccctaattt ccaggtcaag gtgctgtgca
     1261 ttgtggtaat agatgtgaca tgacgtcact tccaaaggac caatgaacat gtctgaccaa
     1321 tttcatataa tgtgaaaacg attttcatag gcagaataaa taacatttaa attaaactgg
     1381 gcatcagcgc aattcaattg gtttggtaat agcaagggaa aatagaatga agtgatctcc
     1441 aaaaaataag tactttttga ctgtaaataa aattgtaagg agtaaaaagt actttttttt
     1501 ctaaaaaaat gtaattaagt aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag
     1561 taaaaatccc caaaaataat acttaagtac agtaatcaag taaaattact caagtacttt
     1621 acacctctgg ttcttgaccc cctaccttca gcaagcccag cagatccact agttctagag
     1681 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     1741 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     1801 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     1861 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     1921 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     1981 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     2041 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     2101 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     2161 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     2221 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     2281 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     2341 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     2401 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     2461 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     2521 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     2581 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     2641 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     2701 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     2761 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     2821 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     2881 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     2941 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     3001 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     3061 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     3121 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     3181 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     3241 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     3301 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     3361 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     3421 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     3481 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     3541 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     3601 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     3661 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     3721 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     3781 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     3841 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     3901 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3961 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     4021 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     4081 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     4141 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     4201 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     4261 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4321 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     4381 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     4441 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     4501 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
     4561 ccaaatagta ggaattaccc acctgtacaa gtgctgaaaa cttggatgaa taagcccgtt
     4621 tgccattttc tactgctatt ttaaatcttt tctgtatttg tctgcatttg tctttaccct
     4681 tgaataaatg ttttagttgt tttttttcaa ttatgactgt gtttaacaga cattattaca
     4741 ataatatgta atagtagtac acactattat tatgtaatag tacttgttga ctgtatttga
     4801 gaactggacc gagtgagtgt tacgtcaccc attcaaaatg acttacttct ggctccaaca
     4861 aaatgaagtt aattcagttg ccatttttca ctgtatggac atcgccgtgt tggagctaga
     4921 cgttgccaat aagcaagaaa gagccgagat gcgtcgagtc tgagtcaccg ttcctatggc
     4981 aacccctcta accaatcaga agtaagcttg ttggaagtcc acagcctacc acttgaaagc
     5041 gggctgcaca aaatctgtca aacgttttga acgttggatg tgagagcaca tacttttatt
     5101 aaggcatctg gttggtcagt ttataatatc aacaacttgg gctacagaaa agaaaagtta
     5161 ttacagaaat tatgattaac aagtacatgt taaataaaga ttttaatatg aatgccacca
     5221 ctggagcatt catgccattt ggagcttctt cctgtttgga tcactagaag gaggaggtca
     5281 ctcattacag ttctcatata cagtcgttgg ttggttggtt ggttggtaga ttgattgatt
     5341 gattgattga ttgattgatt gattgattga ttgattgatt gattgattga ttgattgatt
     5401 ggtagtcaaa ataagaaata atttccacag attcattaca gaaatgatta aatgcataca
     5461 taaaaaactg gggggggggg ggatacaaca acacacttaa gtcacatttg cctacgtaaa
     5521 gaaaagtaaa gaaaatcaat agctatattt tacatctcct ttttttgctg tcttttaatt
     5581 agccttgttt tgctgttatc ttgattgaaa ctgtaacttc ttcacctgct tcttttcttt
     5641 gtaggttttg ccagcccaga aacaggcatg gctgtgcaag gccagagcac catgcacaat
     5701 gcgctgcatg tcttcatgaa cggctcaatg tcctcagtcc agggctcagc caacgacccc
     5761 atcttcctcc ttcaccatgc tttcattgac aggtaacaaa cacgtcatga cattagactg
     5821 cacagttttt gacaaagttc atacaatctg ttgtttatag ctgctacaat tagtgaagtt
     5881 tgtgaatgta cttggatgag cagcgaaaga tcaattgaga tcaattgtta gagtttggtt
     5941 gccctgcaga gcaaagaaca aaaaataatc tggtggcttt actgcgtgag gttattattg
     6001 gtggaataga aacacaaaac ataattgcat ttatttgttt aattttttat cttatcttaa
     6061 ctttcatctt gcatatttgt ttcttacatc atttctagca tctttgagcg ctggctaaga
     6121 actcatcagc ctccccggtc catctaccca cgtaccaatg caccaattgg ccacaatgac
     6181 ggctactaca tggtgccatt ccttcctctt tataggaatg gagactacct cctgtccaac
     6241 aatgctcttg gatacgagta cgcctacctg ttggacccag gtcattgcac aacaccagaa
     6301 atgccctctg atctgcaaaa gacgtgaata tctgttcaga cacccatatc cactctgttc
     6361 cacacaggtc agaggtttgt ccaggagttc ttgacagagg tgtaaaaagt actcaaaaat
     6421 tttactcaag tgaaagtaca agtacttagg gaaaatttta ctcaattaaa agtaaaagta
     6481 tctggctaga atcttacttg agtaaaagta aaaaagtact ccattaaaat tgtacttgag
     6541 tattaaggaa gtaaaagtaa aagcaagaaa gaaaactaga gattcttgtt taagctttta
     6601 atctcaaaaa acattaaatg aaatgcatac aaggttttat cctgctttag aactgtttgt
     6661 atttaattat caaactataa gacagacaat ctaatgccag tacacgctac tcaaagttgt
     6721 aaaacctcag atttaacttc agtagaagct gattctcaaa attgttagtg tcaagcctag
     6781 ctcttttggg gctgaaaagc aatcctgcag tgctgaaaag cctctcacag gcagccgatg
     6841 cgggaagagg tgtattagtc ttgatagaga ggctgcaaat agcaggaaac gtgagcagag
     6901 actccctggt gtctgaaaca caggccagat gggccctcga gtaggtcact gtgacctggt
     6961 cgagtaggtc actgtgacct ggtcgagtag gtcactgtga cctggtcgag taggtcactg
     7021 tgacctggtc gagtaggtca ctgtgacctg gtcgagatat ccagtgacgt aggaagtcca
     7081 tccattcaca gcgcttctat aaaggcgcca gctgaggcgc ctactactcc aaccgcgact
     7141 gcagcgagca actgagaaga ctggatagag ccggcggttc cgcgaacgag cagtgaccgc
     7201 gctcc
//
