LOCUS       sequence-256466-        6551 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral destination vector with C-terminal V5 tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6551)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6551)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSMPUW_6.4_M1-gateway-V5
FEATURES             Location/Qualifiers
     source          1..6551
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             522..702
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    749..873
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    908..1024
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     protein_bind    1093..1217
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1242..1247,1248..1265,1266..1272)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1326..1985
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1392..1411)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2327..2632
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2511..2530
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2673..2797)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2799..2840
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    2977..3565
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3030..3050)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3448..3459)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3755..3935
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        4004..4214
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4073..4208
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4324..4445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4415..4434
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4497..4514)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4668..5256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4748..4767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5508..5547)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(5557..6372)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5557..6372)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6280..6299
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6364..6383
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgggtctctc tggttagacc
      541 agatttgagc ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa
      601 gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga
      661 gatccctcag acccttttag tcagtgtgga aaatctctag cagtggcgcc cgaacaggga
      721 cctgaaagcg aaagggaaac cagaggagct ctctcgacgc aggactcggc ttgctgaagc
      781 gcgcacggca agaggcgagg ggcggcgact gcagagtacg ccaaaatttt gactagcgga
      841 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggaa aatagcggcc
      901 gccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag
      961 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     1021 ttttcggggg atccgcggaa ttcgtccacg gtccaatctc ggtccgaacg agatatcccg
     1081 caaggcgcgc caacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     1141 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     1201 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     1261 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     1321 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1381 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1441 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1501 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     1561 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1621 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1681 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1741 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1801 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1861 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     1921 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     1981 cgtaaagatc tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     2041 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     2101 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     2161 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     2221 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     2281 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     2341 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     2401 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     2461 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     2521 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     2581 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     2641 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     2701 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     2761 tttacgtttc tcgttcagct ttcttgtaca aagtggttgg taagcctatc cctaaccctc
     2821 tcctcggtct cgattctacg tagtaatgaa ccttaggagc caacagctaa gcacacactt
     2881 cgaaagatat atttaaatac acaattaatt aacacaaata cgcgtatctg cagtcgactt
     2941 tgccagaccg gagtccgaac acagctcttc cagcataatc aacctctgga ttacaaaatt
     3001 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3061 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3121 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3181 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3241 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3301 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3361 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
     3421 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3481 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3541 atctcccttt gggccgcctc cccgcttagt actggtacct ttaagaccaa tgacttacaa
     3601 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     3661 ctcccaacga agacaagatt ccggaattta tttgtgaaat ttgtgatgct attgctttat
     3721 ttgtaaaccg gtgcagctgc tttttgcctg tactgggtct ctctggttag accagatctg
     3781 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
     3841 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
     3901 cagacccttt tagtcagtgt ggaaaatctc tagcatctag agtatgcaaa gcatgcatct
     3961 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4021 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4081 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4141 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4201 tggaggccta ggctagagat cataatcagc cataccacat ttgtagaggt tttacttgct
     4261 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     4321 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4381 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4441 tcttatcatg tctgctagct gagcgggctt ttttttctta ggccttcttc cgcttcctcg
     4501 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     4561 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4621 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4681 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4741 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4801 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4861 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4921 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4981 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5041 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5101 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5161 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5221 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5281 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5341 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5401 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5461 gcgatctgtc tatttcgttc atccatagtt gcctgactcc tgcgcagtcc aaaaaaaaag
     5521 gctccaaaag gagcctttaa ttgtatcggt gggcccttag aaaaactcat cgagcatcaa
     5581 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     5641 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     5701 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     5761 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     5821 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     5881 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     5941 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     6001 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     6061 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     6121 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     6181 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     6241 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     6301 atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg tttcccgttg
     6361 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     6421 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggtt
     6481 taaacaaata gtcaaaagcc tccgggcgac tagtcggggt cattagttca tagcccatat
     6541 atggagttcc g
//
