LOCUS       sequence-36064-e        6267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Subach FV, Patterson GH, Renz M, Lippincott-Schwartz J, Verkhusha VV
  TITLE     Bright monomeric photoactivatable red fluorescent protein for 
            two-color super-resolution sptPALM of live cells.
  JOURNAL   J Am Chem Soc. 2010 May 12;132(18):6481-91.
  PUBMED    20394363
REFERENCE   2  (bases 1 to 6267)
  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: "J Am Chem 
            Soc."; date: "2010-05-12"; volume: "132(18)"; pages: "6481-91"
COMMENT     Sequence Label: pVSVG-PATagRFP
FEATURES             Location/Qualifiers
     source          1..6267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(626..673,674..2158)
                     /codon_start=1
                     /product="vesicular stomatitis virus G glycoprotein, 
                     thermosensitive tsO45 mutant"
                     /label=VSV-G tsO45
                     /note="thermosensitive variant of VSV-G Orsay"
                     /translation="MKCLLYLAFLFIGVNCKFTIVFPHNQKGNWKNVPSNYHYCPSSSD
                     LNWHKDLIGTALQVKMPKSHKAIQADGWMCHASKWVTTCDFRWYGPKYITHSIRSFTPS
                     VEQCKESIEQTKQGTWLNPGFPPQSCGYATVTDAEAAIVQVTPHHVLVDEYTGEWVDSQ
                     FINGKCSNDICPTVHNSTTWHSDYKVKGLCDSNLISMDITFSSEDGELSSLGKEGTGFR
                     SNYFAYETGDKACKMQYCKHWGVRLPSGVWFEMADKDLFAAARFPECPEGSSISAPSQT
                     SVDVSLIQDVERILDYSLCQETWSKIRAGLPISPVDLSYLAPKNPGTGPVFTIINGTLK
                     YFETRYIRVDIAAPILSRMVGMISGTTTERELWDDWAPYEDVEIGPNGVLRTSSGYKFP
                     LYMIGHGMLDSDLHLSSKAQVFEHPHIQDAASQLPDDETLFFFGDTGLSKNPIEFVEGW
                     FSSWKSSIASFFFIIGLIIGLFLVLRVGIYLCIKLKHTKKRQIYTDIEMNRLG"
     regulatory      2213..2222
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2219..2932
                     /codon_start=1
                     /product="monomeric photoactivatable red fluorescent 
                     protein (Gunewardene et al., 2011)"
                     /label=PAmKate
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSSTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMKKKTLGWEPSTEKLKPADGGLEGRVD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDRRLEIIKEADKETYWEQHEVAV
                     ARYSDLPSKLGHKLN"
     polyA_signal    3055..3176
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3092..3111)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3146..3165
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3183..3638)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3320..3341)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3532..3551
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3665..3769
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3771..4128
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3771..3962
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3794..3814)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3979..4114
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4163..4957
                     /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             4163..4957
                     /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(4217..4236)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4827..4846
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5145..5164)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5189..5236
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5565..6153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6054..6073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcg ccaccatgaa gtgccttttg tacttagctt ttttattcat
      661 cggggtgaat tgcaagttca ccatagtttt tccacacaac caaaaaggaa actggaaaaa
      721 tgttccttcc aattaccatt attgcccgtc aagctcagat ttaaattggc ataaggactt
      781 aataggcaca gccttacaag tcaaaatgcc caagagtcac aaggctattc aagcagacgg
      841 ttggatgtgt catgcttcca aatgggtcac tacttgtgat ttccgctggt acggaccgaa
      901 gtatataaca cattccatcc gatccttcac tccatctgta gaacaatgca aggaaagcat
      961 tgaacaaacg aaacaaggaa cttggctgaa tccaggcttc cctcctcaaa gttgtggata
     1021 tgcaactgtg acggatgctg aagcagcgat tgtccaggtg actcctcacc atgtgcttgt
     1081 tgatgaatac acaggagaat gggttgattc acagttcatc aacggaaaat gcagcaatga
     1141 catatgcccc actgtccata actccacaac ctggcattcc gactataagg tcaaagggct
     1201 atgtgattct aacctcattt ccatggacat caccttctcc tcagaggacg gagagctatc
     1261 atccctagga aaggagggca cagggttcag aagtaactac tttgcttatg aaaccggaga
     1321 caaggcctgc aaaatgcagt actgcaagca ttggggagtc agactcccat caggtgtctg
     1381 gttcgagatg gctgataagg atctctttgc tgcagccaga ttccctgaat gcccagaagg
     1441 gtcaagtatc tctgctccat ctcagacctc agtggatgta agtctcattc aggacgttga
     1501 gaggatcttg gattattccc tctgccaaga aacctggagc aaaatcagag cgggtcttcc
     1561 catctctcca gtggatctca gctatcttgc tcctaaaaac ccaggaaccg gtcctgtctt
     1621 taccataatc aatggtaccc taaaatactt tgagaccaga tacatcagag tcgatattgc
     1681 tgctccaatc ctctcaagaa tggtcggaat gatcagtgga actaccacag aaagggaact
     1741 gtgggatgac tgggctccat atgaagacgt ggaaattgga cccaatggag ttctgaggac
     1801 cagttcagga tataagtttc ctttatatat gattggacat ggtatgttgg actccgatct
     1861 tcatcttagc tcaaaggctc aggtgtttga acatcctcac attcaagacg ctgcttcgca
     1921 gcttcctgat gatgagactt tatttttttt tggtgatact gggctatcca aaaatccaat
     1981 cgagtttgta gaaggttggt tcagtagttg gaagagctct attgcctctt ttttctttat
     2041 catagggtta atcattggac tattcttggt tctccgagtt ggtatttatc tttgcattaa
     2101 attaaagcac accaagaaaa gacagattta tacagacata gagatgaacc gacttggaaa
     2161 cgaattcggc tccaccggct ccaccggctc caccggcgcg gatccaccgg tcgccaccat
     2221 ggtgtctaag ggcgaagagc tgattaagga gaacatgcac atgaagctgt acatggaggg
     2281 caccgtgaac aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg
     2341 cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat
     2401 cctggctacc agcttcatgt acggcagcag caccttcatc aaccacaccc agggcatccc
     2461 cgacttctgg aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga
     2521 agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta
     2581 caacgtcaag atcagagggg tgaacttccc atccaacggc cctgtgatga agaagaaaac
     2641 actcggctgg gaaccgagca ccgagaaact gaaacccgct gacggcggcc tggaaggcag
     2701 ggtcgacatg gccctgaagc tcgtgggcgg gggccacctg atctgcaact tcaagaccac
     2761 atacagatcc aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggaccg
     2821 cagactggaa atcatcaagg aggccgacaa agagacctac tgggagcagc acgaggtggc
     2881 tgtggccaga tactccgacc tccctagcaa actggggcac aagcttaatt aaagcggccg
     2941 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3001 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3061 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3121 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
     3181 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     3241 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     3301 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     3361 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     3421 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     3481 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     3541 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     3601 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
     3661 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3721 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
     3781 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3841 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3901 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3961 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4021 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     4081 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
     4141 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     4201 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     4261 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     4321 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
     4381 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     4441 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4501 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4561 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4621 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4681 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4741 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4801 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4861 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4921 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4981 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     5041 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     5101 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     5161 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     5221 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     5281 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     5341 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     5401 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5461 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5521 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5581 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5641 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5701 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5761 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5821 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5881 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5941 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6001 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6061 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6121 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6181 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6241 gtggataacc gtattaccgc catgcat
//
