LOCUS       sequence-385268-        4885 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway pME CreERT2 ORF.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4885)
  AUTHORS   Kemmler CL, Moran HR, Murray BF, Scoresby A, Klem JR, Eckert RL, 
            Lepovsky E, Bertho S, Nieuwenhuize S, Burger S, D'Agati G, Betz C, 
            Puller AC, Felker A, Ditrychova K, Botschi S, Affolter M, Rohner N, 
            Lovely CB, Kwan KM, Burger A, Mosimann C
  TITLE     Next-generation plasmids for transgenesis in zebrafish and beyond.
  JOURNAL   Development. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531.
  PUBMED    36975217
REFERENCE   2  (bases 1 to 4885)
  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. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531."
COMMENT     Sequence Label: pME minprom_creERT2
FEATURES             Location/Qualifiers
     source          1..4885
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             299..1327
                     /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             1340..2275
                     /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             1388..2281
                     /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    2327..2448
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2364..2383)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2418..2437
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(2474..2573)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2590..2609)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2591..2609)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2614..2630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2614..2630)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2732..2751)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2743..3552
                     /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(2810..2829)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3699..4287
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4188..4207
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4441..4458
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4617..4644)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4736..4822)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(4871..4885,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(4885,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctccgcggcc gcccccttcc aatctgctca gagaggacag agtgggcagg
      181 agccagcatt gtgtatataa agctgagcag ggtcagttgc ttcttacgtt tgcttctgag
      241 tctgttgtgt tgacttgcaa cctcagaaac agacatccca tgggccgccc ccttcaccat
      301 gtccaattta ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga
      361 tgaggttcgc aagaacctga tggacatgtt cagggatcgc caggcgtttt ctgagcatac
      421 ctggaaaatg cttctgtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg
      481 gaaatggttt cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg
      541 cggtctggca gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg
      601 gtccgggctg ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg
      661 aaaagaaaac gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga
      721 tttcgaccag gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct
      781 ggcatttctg gggattgctt ataacaccct gttacgtata gccgaaattg ccaggatcag
      841 ggttaaagat atctcacgta ctgacggtgg gagaatgtta atccatattg gcagaacgaa
      901 aacgctggtt agcaccgcag gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt
      961 cgagcgatgg atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg
     1021 ggtcagaaaa aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct
     1081 ggaagggatt tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca
     1141 gagatacctg gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg
     1201 cgctggagtt tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt
     1261 catgaactat atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga
     1321 tggcgatctc gagccatctg ctggagacat gagagctgcc aacctttggc caagcccgct
     1381 catgatcaaa cgctctaaga agaacagcct ggccttgtcc ctgacggccg accagatggt
     1441 cagtgccttg ttggatgctg agccccccat actctattcc gagtatgatc ctaccagacc
     1501 cttcagtgaa gcttcgatga tgggcttact gaccaacctg gcagacaggg agctggttca
     1561 catgatcaac tgggcgaaga gggtgccagg ctttgtggat ttgaccctcc atgatcaggt
     1621 ccaccttcta gaatgtgcct ggctagagat cctgatgatt ggtctcgtct ggcgctccat
     1681 ggagcaccca gtgaagctac tgtttgctcc taacttgctc ttggacagga accagggaaa
     1741 atgtgtagag ggcatggtgg agatcttcga catgctgctg gctacatcat ctcggttccg
     1801 catgatgaat ctgcagggag aggagtttgt gtgcctcaaa tctattattt tgcttaattc
     1861 tggagtgtac acatttctgt ccagcaccct gaagtctctg gaagagaagg accatatcca
     1921 ccgagtcctg gacaagatca cagacacttt gatccacctg atggccaagg caggcctgac
     1981 cctgcagcag cagcaccagc ggctggccca gctcctcctc atcctctccc acatcaggca
     2041 catgagtaac aaaggcatgg agcatctgta cagcatgaag tgcaagaacg tggtgcccct
     2101 ctatgacctg ctgctggagg cggcggacgc ccaccgccta catgcgccca ctagccgtgg
     2161 aggggcatcc gtggaggaga cggaccaaag ccacttggcc actgcgggct ctacttcatc
     2221 gcattccttg caaaagtatt acatcacggg ggaggcagag ggtttccctg ccacagcttg
     2281 atgaagatct gagctccctg gcggaattcg gatcttatta aagcagaact tgtttattgc
     2341 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     2401 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgaag
     2461 ggtgggcgcg ccgacccagc tttcttgtac aaagttggca ttataagaaa gcattgctta
     2521 tcaatttgtt gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca
     2581 gctgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg gcagctctgg
     2641 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     2701 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     2761 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2821 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2881 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2941 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     3001 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     3061 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     3121 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     3181 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     3241 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     3301 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     3361 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     3421 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     3481 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     3541 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     3601 tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac
     3661 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3721 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3781 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3841 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3901 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3961 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4021 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4081 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4141 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4201 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4261 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4321 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4381 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4441 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     4501 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     4561 gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa
     4621 ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc
     4681 ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg
     4741 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg
     4801 actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt aacgctagca
     4861 tggatgtttt cccagtcacg acgtt
//
