LOCUS       sequence-201390-        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        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      673..682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             682..705
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             790..846
                     /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             919..939
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1074..1090)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1075..1091)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(1146..1165)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1147..1165)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1439..1560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1476..1495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1530..1549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1567..2022)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1704..1725)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1916..1935
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2049..2153
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2155..2512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2155..2346
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2178..2198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2363..2498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2547..3341
                     /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             2547..3341
                     /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(2601..2620)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3211..3230
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3529..3548)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3573..3620
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3949..4537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4438..4457
                     /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 ccgccaccat ggattacaag gatgacgacg ataagagccc gggcggatcc
      721 aacactgaag ccgccaggcg ttctcgtgcg agaaagttgc aaagaatgaa acaacttgaa
      781 gacaaggttg aagaattgct ttcgaaaaat tatcacttgg aaaatgaggt tgccagatta
      841 aagaaattag ttggcgaacg cgccagccct agcaaccctg gcgccagcaa cggcagcctg
      901 caggaattca gcggccctcc caagaagaag aggaaggtga cctctgatga agtcaggaag
      961 aacctgatgg acatgttcag ggacaggcag gccttctctg aacacacctg gaagatgctc
     1021 ctgtctgtgt gcagatcctg ggctgcctgg tgcaagctga actagaagct tatcgatacc
     1081 gtcgacctcg agggggggcc cggtacctta attaattaag gtaccaggta agtgtaccca
     1141 attcgcccta tagtgagtcg tattacaatt cactcgatcg cccttcccaa cagttgcgca
     1201 gcctgaatgg cgaatggaga tccaattttt aagtgtataa tgtgttaaac tactgattct
     1261 aattgtttgt gtattttaga ttcacagtcc caaggctcat ttcaggcccc tcagtcctca
     1321 cagtctgttc atgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1381 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1441 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1501 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1561 acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1621 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1681 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1741 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1801 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1861 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     1921 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     1981 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag aatcctgagg
     2161 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     2521 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2581 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2641 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2701 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     2761 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2821 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2881 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2941 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3001 ttgtcgatca ggatgatctg gacgaagaac atcaggggct cgcgccagcc gaactgttcg
     3061 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3121 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3181 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagaac
     3241 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3301 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3361 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3421 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3481 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     3541 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3601 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3661 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3721 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3781 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3841 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3901 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4081 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4561 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4621 ttctgtggat aaccgtatta ccgcc
//
