LOCUS       sequence-157297-        7282 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian GFP reporter plasmid, target site for Vika recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7282)
  AUTHORS   Karimova M, Abi-Ghanem J, Berger N, Surendranath V, Pisabarro MT, 
            Buchholz F
  TITLE     Vika/vox, a novel efficient and specific Cre/loxP-like site-specific
            recombination system.
  JOURNAL   Nucleic Acids Res. 2013 Jan;41(2):e37. doi: 10.1093/nar/gks1037. 
            Epub 2012 Nov 9.
  PUBMED    23143104
REFERENCE   2  (bases 1 to 7282)
  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: "Nucleic 
            Acids Res."; date: "2013-01"; volume: "41(2):e37. doi"; pages: " 
            10.1093/nar/gks1037. Epub 2012 Nov 9"
COMMENT     Sequence Label: pRK5-vox-EGFP
FEATURES             Location/Qualifiers
     source          1..7282
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1005..1109
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1111..1468
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1111..1302
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      1319..1454
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1503..2294
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRCQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1557..1576)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2167..2186
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2485..2504)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2529..2576
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             join(2665..2667,2668..2670,2671..3384)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2665..2667,2668..2670,2671..3381)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2710..2731)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2971..2990)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3318..3339
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3391..3420
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     regulatory      3432..3441
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3445..3579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3482..3501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3536..3555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3648..4005
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3648..3839
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     rep_origin      3856..3991
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4025..4042)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4025..4041)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4034..4056)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4254..4709
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4341..4360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4551..4572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4826..4845)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4945..4967
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5005..5023)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5174..5245
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5246..5314,5315..6106)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5464..5483)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6277..6865
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6766..6785
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7019..7036
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7153..7174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7189..7194,7195..7212,7213..7219)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7227..7243
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7232..7254
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7251..7267
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7251..7267
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgacttc gaaagatcta ataggtctga gaacgcccat
      961 tctcagacgt attaagctta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1021 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     1081 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     1141 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     1201 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     1261 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     1321 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     1381 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     1441 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     1501 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     1561 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     1621 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     1681 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     1741 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     1801 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     1861 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     1921 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     1981 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     2041 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     2101 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctgtcagg
     2161 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     2221 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     2281 ttgacgagtt cttctgggcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     2341 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     2401 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     2461 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     2521 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     2581 acgcggggtt cggtcccagg gctggcactc tgtcgataaa taggtctgag aacgcccatt
     2641 ctcagacgta ttctcgaggg atccatggtg agcaagggcg aggagctgtt caccggggtg
     2701 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2761 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2821 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2881 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2941 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     3001 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     3061 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     3121 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     3181 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     3241 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     3301 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     3361 ggcatggacg agctgtacaa gtaagtcgac gagcagaagc tgatcagcga ggaggacctg
     3421 ctgtaagctt ggccgccatg gcccaacttg tttattgcag cttataatgg ttacaaataa
     3481 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3541 ttgtccaaac tcatcaatgt atcttatcat gtctggatcg ggaattaatt cggcgcagca
     3601 ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag
     3661 aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3721 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3781 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     3841 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3901 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3961 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct gttaacagct
     4021 tggcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     4081 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     4141 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     4201 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4261 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4321 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4381 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4441 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4501 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4561 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     4621 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4681 ttttaacaaa atattaacgt ttacaatttt atggtgcact cttgccgatt tcggcctatt
     4741 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt
     4801 ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     4861 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     4921 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     4981 caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5041 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5101 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5161 gataaatgct ttacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5221 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5281 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5341 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5401 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5461 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5521 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5581 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5641 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5701 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5761 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5821 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5881 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5941 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6001 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6061 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6121 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6181 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6241 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6301 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6361 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6421 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6481 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6541 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6601 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6661 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6721 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6781 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6841 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6901 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6961 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     7021 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     7081 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     7141 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     7201 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     7261 ctatgacatg attacgaatt aa
//
