LOCUS       Exported                6361 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  C-terminal part of the split-CreERT2 System.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Tag3B-ERT2CCre
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6361
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        67..370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        371..574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     525..545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        620..638
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      670..679
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             679..708
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             727..1662
                     /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             1777..1833
                     /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             1906..1926
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2790..2806)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2791..2807)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2862..2881)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2863..2881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3155..3276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3192..3211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3246..3265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3283..3738)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3420..3441)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3632..3651
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3765..3869
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3871..4228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3894..3914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4079..4214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4141..4160
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4263..5057
                     /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(4317..4336)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4927..4946
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5245..5264)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5289..5336
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5665..6253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6154..6173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
       61 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      121 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      181 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      241 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      301 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      361 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
      421 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      481 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
      541 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta
      601 gcgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg gagctccacc
      661 gcggtggcgg ccgccatgga gcagaaactc atctctgaag aggatctgag cccgggcgga
      721 tcctctgctg gagacatgag agctgccaac ctttggccaa gcccgctcat gatcaaacgc
      781 tctaagaaga acagcctggc cttgtccctg acggccgacc agatggtcag tgccttgttg
      841 gatgctgagc cccccatact ctattccgag tatgatccta ccagaccctt cagtgaagct
      901 tcgatgatgg gcttactgac caacctggca gacagggagc tggttcacat gatcaactgg
      961 gcgaagaggg tgccaggctt tgtggatttg accctccatg atcaggtcca ccttctagaa
     1021 tgtgcctggc tagagatcct gatgattggt ctcgtctggc gctccatgga gcacccagtg
     1081 aagctactgt ttgctcctaa cttgctcttg gacaggaacc agggaaaatg tgtagagggc
     1141 atggtggaga tcttcgacat gctgctggct acatcatctc ggttccgcat gatgaatctg
     1201 cagggagagg agtttgtgtg cctcaaatct attattttgc ttaattctgg agtgtacaca
     1261 tttctgtcca gcaccctgaa gtctctggaa gagaaggacc atatccaccg agtcctggac
     1321 aagatcacag acactttgat ccacctgatg gccaaggcag gcctgaccct gcagcagcag
     1381 caccagcggc tggcccagct cctcctcatc ctctcccaca tcaggcacat gagtaacaaa
     1441 ggcatggagc atctgtacag catgaagtgc aagaacgtgg tgcccctcta tgacctgctg
     1501 ctggaggcgg cggacgccca ccgcctacat gcgcccacta gccgtggagg ggcatccgtg
     1561 gaggagacgg accaaagcca cttggccact gcgggctcta cttcatcgca ttccttgcaa
     1621 aagtattaca tcacggggga ggcagagggt ttccctgcca cagtcgccag ccctagcaac
     1681 cctggcgcca gcaacggcag cggatccaac actgaagccg ccaggcgttc tcgtgcgaga
     1741 aagttgcaaa gaatgaaaca acttgaagac aaggttgaag aattgctttc gaaaaattat
     1801 cacttggaaa atgaggttgc cagattaaag aaattagttg gcgaacgcgc cagccctagc
     1861 aaccctggcg ccagcaacgg cagcctgcag gaattcagcg gccctcccaa gaagaagagg
     1921 aaggtgaaca ggaaatggtt ccctgctgaa cctgaggatg tgagggacta cctcctgtac
     1981 ctgcaagcca gaggcctggc tgtgaagacc atccaacagc acctgggcca gctcaacatg
     2041 ctgcacagga gatctggcct gcctcgccct tctgactcca atgctgtgtc cctggtgatg
     2101 aggagaatca gaaaggagaa tgtggatgct ggggagagag ccaagcaggc cctggccttt
     2161 gaacgcactg actttgacca agtcagatcc ctgatggaga actctgacag atgccaggac
     2221 atcaggaacc tggccttcct gggcattgcc tacaacaccc tgctgcgcat tgccgaaatt
     2281 gccagaatca gagtgaagga catctcccgc accgatggtg ggagaatgct gatccacatt
     2341 ggcaggacca agaccctggt gtccacagct ggtgtggaga aggccctgtc cctgggggtt
     2401 accaagctgg tggagagatg gatctctgtg tctggtgtgg ctgatgaccc caacaactac
     2461 ctgttctgcc gggtcagaaa gaatggtgtg gctgcccctt ctgccacctc ccaactgtcc
     2521 acccgggccc tggaagggat ctttgaggcc acccaccgcc tgatctatgg tgccaaggat
     2581 gactctgggc agagatacct ggcctggtct ggccactctg ccagagtggg tgctgccagg
     2641 gacatggcca gggctggtgt gtccatccct gaaatcatgc aggctggtgg ctggaccaat
     2701 gtgaacattg tgatgaacta catcagaaac ctggactctg agactggggc catggtgagg
     2761 ctgctcgagg atggggacta gaagcttatc gataccgtcg acctcgaggg ggggcccggt
     2821 accttaatta attaaggtac caggtaagtg tacccaattc gccctatagt gagtcgtatt
     2881 acaattcact cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggagatcca
     2941 atttttaagt gtataatgtg ttaaactact gattctaatt gtttgtgtat tttagattca
     3001 cagtcccaag gctcatttca ggcccctcag tcctcacagt ctgttcatga tcataatcag
     3061 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3121 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3181 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3241 tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta agcgttaata
     3301 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3361 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3421 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3481 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3541 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3601 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3661 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3721 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3781 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3841 tgcttcaata atattgaaaa aggaagaatc ctgaggcgga aagaaccagc tgtggaatgt
     3901 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3961 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4021 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4081 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4141 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4201 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4261 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4321 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4381 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4441 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4501 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4561 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4621 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4681 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4741 aagaacatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4801 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4861 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4921 acatagcgtt ggctacccgt gatattgctg aagaacttgg cggcgaatgg gctgaccgct
     4981 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5041 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5101 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5161 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5221 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5281 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5341 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5401 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5461 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5521 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5581 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6301 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6361 c
//
