LOCUS       sequence-206083-        6361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C-terminal part of the split-CreERT2 System.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6361)
  AUTHORS   Hirrlinger J, Requardt RP, Winkler U, Wilhelm F, Schulze C, 
            Hirrlinger PG
  TITLE     Split-CreERT2: temporal control of DNA recombination mediated by 
            split-Cre protein fragment complementation.
  JOURNAL   PLoS One. 2009 Dec 16;4(12):e8354. doi: 
            10.1371/journal.pone.0008354.
  PUBMED    20016782
REFERENCE   2  (bases 1 to 6361)
  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: "PLoS One.";
            date: "2009-12-16"; volume: "4(12):e8354. doi"; pages: " 
            10.1371/journal.pone.0008354"
COMMENT     Sequence Label: pCMV-Tag3B-ERT2CCre
FEATURES             Location/Qualifiers
     source          1..6361
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        69..372
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        373..576
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     527..547
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        622..640
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      672..681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             681..710
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             729..1664
                     /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             777..1667
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     CDS             1779..1835
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     CDS             1908..1928
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2792..2808)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2793..2809)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2864..2883)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2865..2883)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3157..3278
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3194..3213)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3248..3267
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3285..3740)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3422..3443)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3634..3653
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3767..3871
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3873..4230
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3873..4064
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3896..3916)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4081..4216
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4265..5059
                     /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             4265..5059
                     /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(4319..4338)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4929..4948
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5247..5266)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5291..5338
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5667..6255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6156..6175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
       61 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
      121 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
      181 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
      241 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
      301 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
      361 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      421 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      481 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      541 aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc
      601 tagcgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca
      661 ccgcggtggc ggccgccatg gagcagaaac tcatctctga agaggatctg agcccgggcg
      721 gatcctctgc tggagacatg agagctgcca acctttggcc aagcccgctc atgatcaaac
      781 gctctaagaa gaacagcctg gccttgtccc tgacggccga ccagatggtc agtgccttgt
      841 tggatgctga gccccccata ctctattccg agtatgatcc taccagaccc ttcagtgaag
      901 cttcgatgat gggcttactg accaacctgg cagacaggga gctggttcac atgatcaact
      961 gggcgaagag ggtgccaggc tttgtggatt tgaccctcca tgatcaggtc caccttctag
     1021 aatgtgcctg gctagagatc ctgatgattg gtctcgtctg gcgctccatg gagcacccag
     1081 tgaagctact gtttgctcct aacttgctct tggacaggaa ccagggaaaa tgtgtagagg
     1141 gcatggtgga gatcttcgac atgctgctgg ctacatcatc tcggttccgc atgatgaatc
     1201 tgcagggaga ggagtttgtg tgcctcaaat ctattatttt gcttaattct ggagtgtaca
     1261 catttctgtc cagcaccctg aagtctctgg aagagaagga ccatatccac cgagtcctgg
     1321 acaagatcac agacactttg atccacctga tggccaaggc aggcctgacc ctgcagcagc
     1381 agcaccagcg gctggcccag ctcctcctca tcctctccca catcaggcac atgagtaaca
     1441 aaggcatgga gcatctgtac agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc
     1501 tgctggaggc ggcggacgcc caccgcctac atgcgcccac tagccgtgga ggggcatccg
     1561 tggaggagac ggaccaaagc cacttggcca ctgcgggctc tacttcatcg cattccttgc
     1621 aaaagtatta catcacgggg gaggcagagg gtttccctgc cacagtcgcc agccctagca
     1681 accctggcgc cagcaacggc agcggatcca acactgaagc cgccaggcgt tctcgtgcga
     1741 gaaagttgca aagaatgaaa caacttgaag acaaggttga agaattgctt tcgaaaaatt
     1801 atcacttgga aaatgaggtt gccagattaa agaaattagt tggcgaacgc gccagcccta
     1861 gcaaccctgg cgccagcaac ggcagcctgc aggaattcag cggccctccc aagaagaaga
     1921 ggaaggtgaa caggaaatgg ttccctgctg aacctgagga tgtgagggac tacctcctgt
     1981 acctgcaagc cagaggcctg gctgtgaaga ccatccaaca gcacctgggc cagctcaaca
     2041 tgctgcacag gagatctggc ctgcctcgcc cttctgactc caatgctgtg tccctggtga
     2101 tgaggagaat cagaaaggag aatgtggatg ctggggagag agccaagcag gccctggcct
     2161 ttgaacgcac tgactttgac caagtcagat ccctgatgga gaactctgac agatgccagg
     2221 acatcaggaa cctggccttc ctgggcattg cctacaacac cctgctgcgc attgccgaaa
     2281 ttgccagaat cagagtgaag gacatctccc gcaccgatgg tgggagaatg ctgatccaca
     2341 ttggcaggac caagaccctg gtgtccacag ctggtgtgga gaaggccctg tccctggggg
     2401 ttaccaagct ggtggagaga tggatctctg tgtctggtgt ggctgatgac cccaacaact
     2461 acctgttctg ccgggtcaga aagaatggtg tggctgcccc ttctgccacc tcccaactgt
     2521 ccacccgggc cctggaaggg atctttgagg ccacccaccg cctgatctat ggtgccaagg
     2581 atgactctgg gcagagatac ctggcctggt ctggccactc tgccagagtg ggtgctgcca
     2641 gggacatggc cagggctggt gtgtccatcc ctgaaatcat gcaggctggt ggctggacca
     2701 atgtgaacat tgtgatgaac tacatcagaa acctggactc tgagactggg gccatggtga
     2761 ggctgctcga ggatggggac tagaagctta tcgataccgt cgacctcgag ggggggcccg
     2821 gtaccttaat taattaaggt accaggtaag tgtacccaat tcgccctata gtgagtcgta
     2881 ttacaattca ctcgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggagatc
     2941 caatttttaa gtgtataatg tgttaaacta ctgattctaa ttgtttgtgt attttagatt
     3001 cacagtccca aggctcattt caggcccctc agtcctcaca gtctgttcat gatcataatc
     3061 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3121 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3181 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3241 tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg taagcgttaa
     3301 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3361 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3421 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3481 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3541 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3601 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3661 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3721 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3781 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3841 aatgcttcaa taatattgaa aaaggaagaa tcctgaggcg gaaagaacca gctgtggaat
     3901 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3961 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4021 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4081 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4141 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4201 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4261 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4321 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4381 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4441 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4501 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4561 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4621 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4681 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4741 cgaagaacat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4801 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4861 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4921 ggacatagcg ttggctaccc gtgatattgc tgaagaactt ggcggcgaat gggctgaccg
     4981 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5041 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5101 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5161 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5221 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5281 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5341 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5401 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5461 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5521 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5581 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5641 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5701 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5761 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5821 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5881 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5941 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6001 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6061 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6121 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6181 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6241 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6301 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6361 g
//
