LOCUS       Exported                5599 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  N-terminal part of the split-CreERT2 System.
ACCESSION   .
VERSION     .
KEYWORDS    pCMV-Tag2B-NCreERT2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5599)
  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 5599)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5599
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        63..366
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        367..570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     521..541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        616..634
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      669..678
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             678..701
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             786..842
                     /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             915..935
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1068..2003
                     /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"
     primer_bind     complement(2024..2040)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2025..2041)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2096..2115)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2097..2115)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2389..2510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2426..2445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2480..2499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2517..2972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2654..2675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2866..2885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2999..3103
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3105..3462
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3128..3148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3313..3448
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3375..3394
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3497..4291
                     /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(3551..3570)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4161..4180
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4479..4498)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4523..4570
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4899..5487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5388..5407
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
       61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      541 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcga
      601 ttacgccaag ctcgaaatta accctcacta aagggaacaa aagctggagc tccaccgcgg
      661 tggcggccgc caccatggat tacaaggatg acgacgataa gagcccgggc ggatccaaca
      721 ctgaagccgc caggcgttct cgtgcgagaa agttgcaaag aatgaaacaa cttgaagaca
      781 aggttgaaga attgctttcg aaaaattatc acttggaaaa tgaggttgcc agattaaaga
      841 aattagttgg cgaacgcgcc agccctagca accctggcgc cagcaacggc agcctgcagg
      901 aattcagcgg ccctcccaag aagaagagga aggtgacctc tgatgaagtc aggaagaacc
      961 tgatggacat gttcagggac aggcaggcct tctctgaaca cacctggaag atgctcctgt
     1021 ctgtgtgcag atcctgggct gcctggtgca agctgaacac tagttctgct ggagacatga
     1081 gagctgccaa cctttggcca agcccgctca tgatcaaacg ctctaagaag aacagcctgg
     1141 ccttgtccct gacggccgac cagatggtca gtgccttgtt ggatgctgag ccccccatac
     1201 tctattccga gtatgatcct accagaccct tcagtgaagc ttcgatgatg ggcttactga
     1261 ccaacctggc agacagggag ctggttcaca tgatcaactg ggcgaagagg gtgccaggct
     1321 ttgtggattt gaccctccat gatcaggtcc accttctaga atgtgcctgg ctagagatcc
     1381 tgatgattgg tctcgtctgg cgctccatgg agcacccagt gaagctactg tttgctccta
     1441 acttgctctt ggacaggaac cagggaaaat gtgtagaggg catggtggag atcttcgaca
     1501 tgctgctggc tacatcatct cggttccgca tgatgaatct gcagggagag gagtttgtgt
     1561 gcctcaaatc tattattttg cttaattctg gagtgtacac atttctgtcc agcaccctga
     1621 agtctctgga agagaaggac catatccacc gagtcctgga caagatcaca gacactttga
     1681 tccacctgat ggccaaggca ggcctgaccc tgcagcagca gcaccagcgg ctggcccagc
     1741 tcctcctcat cctctcccac atcaggcaca tgagtaacaa aggcatggag catctgtaca
     1801 gcatgaagtg caagaacgtg gtgcccctct atgacctgct gctggaggcg gcggacgccc
     1861 accgcctaca tgcgcccact agccgtggag gggcatccgt ggaggagacg gaccaaagcc
     1921 acttggccac tgcgggctct acttcatcgc attccttgca aaagtattac atcacggggg
     1981 aggcagaggg tttccctgcc acagtctgaa ctagtaagct tatcgatacc gtcgacctcg
     2041 agggggggcc cggtacctta attaattaag gtaccaggta agtgtaccca attcgcccta
     2101 tagtgagtcg tattacaatt cactcgatcg cccttcccaa cagttgcgca gcctgaatgg
     2161 cgaatggaga tccaattttt aagtgtataa tgtgttaaac tactgattct aattgtttgt
     2221 gtattttaga ttcacagtcc caaggctcat ttcaggcccc tcagtcctca cagtctgttc
     2281 atgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2341 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2401 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2461 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     2521 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2581 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2641 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2701 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2761 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2821 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2881 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2941 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3001 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3061 ataaccctga taaatgcttc aataatattg aaaaaggaag aatcctgagg cggaaagaac
     3121 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3181 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3301 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3361 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3421 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3481 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3541 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3601 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3661 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3721 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3781 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3841 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3901 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3961 ggatgatctg gacgaagaac atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4021 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4081 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4141 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagaac ttggcggcga
     4201 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4261 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4321 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4381 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4441 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4501 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4561 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4621 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4681 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4741 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4801 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4861 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4921 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4981 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5041 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5101 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5161 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5221 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5281 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5341 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5401 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5461 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5521 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5581 aaccgtatta ccgccatgc
//
