LOCUS       sequence-275157-        4739 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4739)
  AUTHORS   Mosimann C, Kaufman CK, Li P, Pugach EK, Tamplin OJ, Zon LI
  TITLE     Ubiquitous transgene expression and Cre-based recombination driven 
            by the ubiquitin promoter in zebrafish.
  JOURNAL   Development. 2011 Jan . 138(1):169-77.
  PUBMED    21138979
REFERENCE   2  (bases 1 to 4739)
  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"; journalName: 
            "Development. 2011 Jan . 138(1):169-77."
COMMENT     Sequence Label: pENTR/D_creERt2
FEATURES             Location/Qualifiers
     source          1..4739
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      27..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     769..786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(945..972)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1064..1150)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1199..1221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1213..1230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1214..1230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1246..1345
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1366..2394
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     CDS             2407..3342
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2455..3348
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     polyA_signal    3394..3515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3431..3450)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3485..3504
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(3541..3640)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3657..3676)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3658..3676)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3681..3697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3681..3697)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3799..3818)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3810..4619
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(3877..3896)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
       61 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
      121 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
      181 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      781 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
      841 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      901 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
      961 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     1021 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     1081 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     1141 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     1201 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta
     1261 ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg
     1321 ccaactttgt acaaaaaagc aggctccgcg gccgccccct tcaccatgtc caatttactg
     1381 accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa cgagtgatga ggttcgcaag
     1441 aacctgatgg acatgttcag ggatcgccag gcgttttctg agcatacctg gaaaatgctt
     1501 ctgtccgttt gccggtcgtg ggcggcatgg tgcaagttga ataaccggaa atggtttccc
     1561 gcagaacctg aagatgttcg cgattatctt ctatatcttc aggcgcgcgg tctggcagta
     1621 aaaactatcc agcaacattt gggccagcta aacatgcttc atcgtcggtc cgggctgcca
     1681 cgaccaagtg acagcaatgc tgtttcactg gttatgcggc ggatccgaaa agaaaacgtt
     1741 gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac gcactgattt cgaccaggtt
     1801 cgttcactca tggaaaatag cgatcgctgc caggatatac gtaatctggc atttctgggg
     1861 attgcttata acaccctgtt acgtatagcc gaaattgcca ggatcagggt taaagatatc
     1921 tcacgtactg acggtgggag aatgttaatc catattggca gaacgaaaac gctggttagc
     1981 accgcaggtg tagagaaggc acttagcctg ggggtaacta aactggtcga gcgatggatt
     2041 tccgtctctg gtgtagctga tgatccgaat aactacctgt tttgccgggt cagaaaaaat
     2101 ggtgttgccg cgccatctgc caccagccag ctatcaactc gcgccctgga agggattttt
     2161 gaagcaactc atcgattgat ttacggcgct aaggatgact ctggtcagag atacctggcc
     2221 tggtctggac acagtgcccg tgtcggagcc gcgcgagata tggcccgcgc tggagtttca
     2281 ataccggaga tcatgcaagc tggtggctgg accaatgtaa atattgtcat gaactatatc
     2341 cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc tggaagatgg cgatctcgag
     2401 ccatctgctg gagacatgag agctgccaac ctttggccaa gcccgctcat gatcaaacgc
     2461 tctaagaaga acagcctggc cttgtccctg acggccgacc agatggtcag tgccttgttg
     2521 gatgctgagc cccccatact ctattccgag tatgatccta ccagaccctt cagtgaagct
     2581 tcgatgatgg gcttactgac caacctggca gacagggagc tggttcacat gatcaactgg
     2641 gcgaagaggg tgccaggctt tgtggatttg accctccatg atcaggtcca ccttctagaa
     2701 tgtgcctggc tagagatcct gatgattggt ctcgtctggc gctccatgga gcacccagtg
     2761 aagctactgt ttgctcctaa cttgctcttg gacaggaacc agggaaaatg tgtagagggc
     2821 atggtggaga tcttcgacat gctgctggct acatcatctc ggttccgcat gatgaatctg
     2881 cagggagagg agtttgtgtg cctcaaatct attattttgc ttaattctgg agtgtacaca
     2941 tttctgtcca gcaccctgaa gtctctggaa gagaaggacc atatccaccg agtcctggac
     3001 aagatcacag acactttgat ccacctgatg gccaaggcag gcctgaccct gcagcagcag
     3061 caccagcggc tggcccagct cctcctcatc ctctcccaca tcaggcacat gagtaacaaa
     3121 ggcatggagc atctgtacag catgaagtgc aagaacgtgg tgcccctcta tgacctgctg
     3181 ctggaggcgg cggacgccca ccgcctacat gcgcccacta gccgtggagg ggcatccgtg
     3241 gaggagacgg accaaagcca cttggccact gcgggctcta cttcatcgca ttccttgcaa
     3301 aagtattaca tcacggggga ggcagagggt ttccctgcca cagcttgatg aagatctgag
     3361 ctccctggcg gaattcggat cttattaaag cagaacttgt ttattgcagc ttataatggt
     3421 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3481 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgaagggt gggcgcgccg
     3541 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     3601 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     3661 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     3721 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     3781 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     3841 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     3901 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     3961 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     4021 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     4081 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     4141 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     4201 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     4261 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     4321 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     4381 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     4441 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     4501 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     4561 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     4621 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     4681 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagac
//
