LOCUS       sequence-270525-        8131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral destination vector with Ef1a promoter, N-terminal HA tag,
            and SV40 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8131)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8131)
  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            "Unpublished"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_6.10_M3_Ef1a-HA-gateway-SV40
FEATURES             Location/Qualifiers
     source          1..8131
                     /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"
     promoter        1158..2336
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1389..2327
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2284..2304
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             2400..2426
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    2427..2551
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2576..2581,2582..2599,2600..2606)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2660..3319
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2726..2745)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3661..3966
                     /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     3845..3864
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4007..4131)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        4224..4434
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4246..4442
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4293..4428
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4557..5145
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4610..4630)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5028..5039)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5335..5515
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        5584..5794
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5653..5788
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5904..6025
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5995..6014
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6077..6094)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6248..6836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6328..6347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(7088..7127)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(7137..7952)
                     /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(7137..7952)
                     /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     7860..7879
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7944..7963
                     /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 agatatctgc
     1081 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
     1141 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
     1201 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
     1261 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
     1321 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
     1381 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
     1441 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
     1501 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
     1561 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     1621 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     1681 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     1741 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1801 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1861 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1921 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1981 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     2041 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     2101 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     2161 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     2221 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     2281 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggcg
     2341 cgcctatcga aattcactat agggagaccc aagctggcta gttaagcttc gccaccatgt
     2401 acccatacga tgttccagat tacgctacaa gtttgtacaa aaaagctgaa cgagaaacgt
     2461 aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag actacataat
     2521 actgtaaaac acaacatatc cagtcactat ggcggccgca ttaggcaccc caggctttac
     2581 actttatgct tccggctcgt ataatgtgtg gattttgagt taggatccgg cgagattttc
     2641 aggagctaag gaagctaaaa tggagaaaaa aatcactgga tataccaccg ttgatatatc
     2701 ccaatggcat cgtaaagaac attttgaggc atttcagtca gttgctcaat gtacctataa
     2761 ccagaccgtt cagctggata ttacggcctt tttaaagacc gtaaagaaaa ataagcacaa
     2821 gttttatccg gcctttattc acattcttgc ccgcctgatg aatgctcatc cggaattccg
     2881 tatggcaatg aaagacggtg agctggtgat atgggatagt gttcaccctt gttacaccgt
     2941 tttccatgag caaactgaaa cgttttcatc gctctggagt gaataccacg acgatttccg
     3001 gcagtttcta cacatatatt cgcaagatgt ggcgtgttac ggtgaaaacc tggcctattt
     3061 ccctaaaggg tttattgaga atatgttttt cgtctcagcc aatccctggg tgagtttcac
     3121 cagttttgat ttaaacgtgg ccaatatgga caacttcttc gcccccgttt tcaccatggg
     3181 caaatattat acgcaaggcg acaaggtgct gatgccgctg gcgattcagg ttcatcatgc
     3241 cgtctgtgat ggcttccatg tcggcagaat gcttaatgaa ttacaacagt actgcgatga
     3301 gtggcagggc ggggcgtaaa cgcgtggatc cggcttacta aaagccagat aacagtatgc
     3361 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
     3421 gtcaaaaaga ggtgtgctat gaagcagcgt attacagtga cagttgacag cgacagctat
     3481 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     3541 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     3601 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg gactggtgaa
     3661 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     3721 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     3781 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     3841 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     3901 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     3961 atataaatgt caggctccgt tatacacagc cagtctgcag gtcgaccata gtgactggat
     4021 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     4081 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg tagtaatgac
     4141 cttaggagcc aacagctaag cacacacttc gaatgcatct caattagtca gcaaccaggt
     4201 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcgt ctcaattagt
     4261 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4321 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4381 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4441 aaaagctttc tagaggtacc accggagctt atttaaatac acaattaatt aacacaaata
     4501 cgcgtatctg cagtcgactt tgccagaccg gagtccgaac acagctcttc cagcataatc
     4561 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     4621 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     4681 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4741 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4801 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4861 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4921 gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct
     4981 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     5041 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     5101 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcttagt actggtacct
     5161 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     5221 ggactggaag ggctaattca ctcccaacga agacaagatt ccggaattta tttgtgaaat
     5281 ttgtgatgct attgctttat ttgtaaaccg gtgcagctgc tttttgcctg tactgggtct
     5341 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     5401 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     5461 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcatctag
     5521 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     5581 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     5641 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     5701 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     5761 aagtagtgag gaggcttttt tggaggccta ggctagagat cataatcagc cataccacat
     5821 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5881 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5941 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6001 tgtccaaact catcaatgta tcttatcatg tctgctagct gagcgggctt ttttttctta
     6061 ggccttcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6121 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6181 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6241 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6301 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6361 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6421 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6481 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6541 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6601 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6661 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6721 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6781 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6961 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7021 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7081 tgcgcagtcc aaaaaaaaag gctccaaaag gagcctttaa ttgtatcggt gggcccttag
     7141 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     7201 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     7261 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     7321 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     7381 tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca
     7441 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     7501 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     7561 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     7621 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     7681 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     7741 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     7801 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     7861 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc
     7921 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     7981 gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga
     8041 ttttgagaca caacgtggtt taaacaaata gtcaaaagcc tccgggcgac tagtcggggt
     8101 cattagttca tagcccatat atggagttcc g
//
