LOCUS       sequence-207091-        5818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High copy number plasmid containing iCreERT2-FFrt-KanR-FFrt insert.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  AUTHORS   Korecki AJ, Hickmott JW, Lam SL, Dreolini L, Mathelier A, Baker O, 
            Kuehne C, Bonaguro RJ, Smith J, Tan CV, Zhou M, Goldowitz D, 
            Deussing JM, Stewart AF, Wasserman WW, Holt RA, Simpson EM
  TITLE     Twenty-Seven Tamoxifen-Inducible iCre-Driver Mouse Strains for Eye 
            and Brain; Including Seventeen Carrying a New Inducible-First 
            Constitutive-Ready Allele.
  JOURNAL   Genetics. 2019 Feb 14. pii: genetics.119.301984. doi: 
            10.1534/genetics.119.301984.
  PUBMED    30765420
REFERENCE   2  (bases 1 to 5818)
  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: "Genetics. 
            2019 Feb 14. pii: genetics.119.301984. doi: 
            10.1534/genetics.119.301984."
COMMENT     Sequence Label: pEMS1725
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1244..1277
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             2329..3264
                     /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             2377..3270
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     protein_bind    3271..3318
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3457..4251
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3457..4251
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3511..3530)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4121..4140
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     protein_bind    4252..4299
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    complement(4318..4351)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(4821..4838)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4821..4837)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4936..4953)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5107..5695)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5187..5206)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5817..5818,1..790,791..859))
                     /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"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttat tcgactctat agtgaagttc ctattctcta
     1261 gaaagtatag gaacttctga agtggggatg tccaacctgc tgactgtgca ccaaaacctg
     1321 cctgccctcc ctgtggatgc cacctctgat gaagtcagga agaacctgat ggacatgttc
     1381 agggacaggc aggccttctc tgaacacacc tggaagatgc tcctgtctgt gtgcagatcc
     1441 tgggctgcct ggtgcaagct gaacaacagg aaatggttcc ctgctgaacc tgaggatgtg
     1501 agggactacc tcctgtacct gcaagccaga ggcctggctg tgaagaccat ccaacagcac
     1561 ctgggccagc tcaacatgct gcacaggaga tctggcctgc ctcgcccttc tgactccaat
     1621 gctgtgtccc tggtgatgag gagaatcaga aaggagaatg tggatgctgg ggagagagcc
     1681 aagcaggccc tggcctttga acgcactgac tttgaccaag tcagatccct gatggagaac
     1741 tctgacagat gccaggacat caggaacctg gccttcctgg gcattgccta caacaccctg
     1801 ctgcgcattg ccgaaattgc cagaatcaga gtgaaggaca tctcccgcac cgatggtggg
     1861 agaatgctga tccacattgg caggaccaag accctggtgt ccacagctgg tgtggagaag
     1921 gccctgtccc tgggggttac caagctggtg gagagatgga tctctgtgtc tggtgtggct
     1981 gatgacccca acaactacct gttctgccgg gtcagaaaga atggtgtggc tgccccttct
     2041 gccacctccc aactgtccac ccgggccctg gaagggatct ttgaggccac ccaccgcctg
     2101 atctatggtg ccaaggatga ctctgggcag agatacctgg cctggtctgg ccactctgcc
     2161 agagtgggtg ctgccaggga catggccagg gctggtgtgt ccatccctga aatcatgcag
     2221 gctggtggct ggaccaatgt gaacatagtg atgaactaca tcagaaacct ggactctgag
     2281 actggggcca tggtgaggct gctcgaggat ggggacctcg agccatctgc tggagacatg
     2341 agagctgcca acctttggcc aagcccgctc atgatcaaac gctctaagaa gaacagcctg
     2401 gccttgtccc tgacggccga ccagatggtc agtgccttgt tggatgctga gccccccata
     2461 ctctattccg agtatgatcc taccagaccc ttcagtgaag cttcgatgat gggcttactg
     2521 accaacctgg cagacaggga gctggttcac atgatcaact gggcgaagag ggtgccaggc
     2581 tttgtggatt tgaccctcca tgatcaggtc caccttctag aatgtgcctg gctagagatc
     2641 ctgatgattg gtctcgtctg gcgctccatg gagcacccag tgaagctact gtttgctcct
     2701 aacttgctct tggacaggaa ccagggaaaa tgtgtagagg gcatggtgga gatcttcgac
     2761 atgctgctgg ctacatcatc tcggttccgc atgatgaatc tgcagggaga ggagtttgtg
     2821 tgcctcaaat ctattatttt gcttaattct ggagtgtaca catttctgtc cagcaccctg
     2881 aagtctctgg aagagaagga ccatatccac cgagtcctgg acaagatcac agacactttg
     2941 atccacctga tggccaaggc aggcctgacc ctgcagcagc agcaccagcg gctggcccag
     3001 ctcctcctca tcctctccca catcaggcac atgagtaaca aaggcatgga gcatctgtac
     3061 agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc tgctggaggc ggcggacgcc
     3121 caccgcctac atgcgcccac tagccgtgga ggggcatccg tggaggagac ggaccaaagc
     3181 cacttggcca ctgcgggctc tacttcatcg cattccttgc aaaagtatta catcacgggg
     3241 gaggcagagg gtttccctgc cacagcttga gaagttccta ttccgaagtt cctattctct
     3301 agaaagtata ggaacttccc ggaattgcca gctggggcgc cctctggtaa ggttgggaag
     3361 ccctgcaaag taaactggat ggctttctcg ccgccaagga tctgatggcg caggggatca
     3421 agctctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3481 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3541 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3601 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3661 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3721 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3781 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3841 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     3901 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     3961 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4021 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4081 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4141 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4201 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agaagttcct
     4261 attccgaagt tcctattctc tagaaagtat aggaacttca attaattaaa atgaagtgaa
     4321 gttcctatac tttctagaga ataggaactt ctatagtgag tcgaataagg gcgacacaaa
     4381 atttattcta aatgcataat aaatactgat aacatcttat agtttgtatt atattttgta
     4441 ttatcgttga catgtataat tttgatatca aaaactgatt ttccctttat tattttcgag
     4501 atttattttc ttaattctct ttaacaaact agaaatattg tatatacaaa aaatcataaa
     4561 taatagatga atagtttaat tataggtgtt catcaatcga aaaagcaacg tatcttattt
     4621 aaagtgcgtt gcttttttct catttataag gttaaataat tctcatatat caagcaaagt
     4681 gacaggcgcc cttaaatatt ctgacaaatg ctctttccct aaactccccc cataaaaaaa
     4741 cccgccgaag cgggttttta cgttatttgc ggattaacga ttactcgtta tcagaaccgc
     4801 ccagggggcc cgagcttaag actggccgtc gttttacaac acagaaagag tttgtagaaa
     4861 cgcaaaaagg ccatccgtca ggggccttct gcttagtttg atgcctggca gttccctact
     4921 ctcgccttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4981 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5041 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5101 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5161 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5221 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5281 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5341 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5401 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5461 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5521 gtgggctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5581 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5641 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt
     5761 taagggattt tggtcatgag cttgcgccgt cccgtcaagt cagcgtaatg ctctgctt
//
