LOCUS       sequence-206064-        4645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal part of the split-Cre System.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4645)
  AUTHORS   Hirrlinger J, Scheller A, Hirrlinger PG, Kellert B, Tang W, Wehr MC,
            Goebbels S, Reichenbach A, Sprengel R, Rossner MJ, Kirchhoff F
  TITLE     Split-cre complementation indicates coincident activity of different
            genes in vivo.
  JOURNAL   PLoS One. 2009;4(1):e4286. doi: 10.1371/journal.pone.0004286. Epub 
            2009 Jan 27.
  PUBMED    19172189
REFERENCE   2  (bases 1 to 4645)
  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"; volume: "4(1):e4286. doi"; pages: " 
            10.1371/journal.pone.0004286. Epub 2009 Jan 27"
COMMENT     Sequence Label: pCMV-Tag2B-NCre
FEATURES             Location/Qualifiers
     source          1..4645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        612..630
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             674..697
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             782..838
                     /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             911..931
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1066..1082)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1067..1083)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1138..1157)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1139..1157)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1431..1552
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1468..1487)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1522..1541
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1559..2014)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1696..1717)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1908..1927
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2041..2145
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2147..2504
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2147..2338
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2170..2190)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2355..2490
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2539..3333
                     /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             2539..3333
                     /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(2593..2612)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3203..3222
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3521..3540)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3565..3612
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3941..4529
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4430..4449
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgattac
      601 gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc
      661 ggccgccacc atggattaca aggatgacga cgataagagc ccgggcggat ccaacactga
      721 agccgccagg cgttctcgtg cgagaaagtt gcaaagaatg aaacaacttg aagacaaggt
      781 tgaagaattg ctttcgaaaa attatcactt ggaaaatgag gttgccagat taaagaaatt
      841 agttggcgaa cgcgccagcc ctagcaaccc tggcgccagc aacggcagcc tgcaggaatt
      901 cagcggccct cccaagaaga agaggaaggt gacctctgat gaagtcagga agaacctgat
      961 ggacatgttc agggacaggc aggccttctc tgaacacacc tggaagatgc tcctgtctgt
     1021 gtgcagatcc tgggctgcct ggtgcaagct gaactagaag cttatcgata ccgtcgacct
     1081 cgaggggggg cccggtacct taattaatta aggtaccagg taagtgtacc caattcgccc
     1141 tatagtgagt cgtattacaa ttcactcgat cgcccttccc aacagttgcg cagcctgaat
     1201 ggcgaatgga gatccaattt ttaagtgtat aatgtgttaa actactgatt ctaattgttt
     1261 gtgtatttta gattcacagt cccaaggctc atttcaggcc cctcagtcct cacagtctgt
     1321 tcatgatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     1381 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     1441 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1501 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa
     1561 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     1621 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     1681 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     1741 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     1801 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     1861 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     1921 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     1981 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     2041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     2101 caataaccct gataaatgct tcaataatat tgaaaaagga agaatcctga ggcggaaaga
     2161 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     2221 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     2281 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     2341 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     2401 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     2461 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     2521 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2581 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2641 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2701 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     2761 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2821 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2881 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2941 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3001 caggatgatc tggacgaaga acatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3061 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3121 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3181 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga acttggcggc
     3241 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3301 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3361 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3421 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3481 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     3541 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     3601 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3661 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3721 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     3781 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3841 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3901 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3961 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4021 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4081 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4141 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4201 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4261 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4321 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4381 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4441 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4501 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4561 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4621 ataaccgtat taccgccatg catta
//
