LOCUS       sequence-256451-        8080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral destination vector with Ef1a promoter, C-terminal HA tag,
            and SV40 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8080)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8080)
  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            "Unpublished"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_6.11_M3_Ef1a-gateway-HA-SV40
FEATURES             Location/Qualifiers
     source          1..8080
                     /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"
     protein_bind    2346..2470
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2495..2500,2501..2518,2519..2525)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2579..3238
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2645..2664)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3580..3885
                     /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     3764..3783
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3926..4050)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4052..4078
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        4173..4383
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4195..4391
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4242..4377
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4506..5094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4559..4579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4977..4988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5284..5464
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        5533..5743
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5602..5737
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5853..5974
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5944..5963
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6026..6043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6197..6785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6277..6296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(7037..7076)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(7086..7901)
                     /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(7086..7901)
                     /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     7809..7828
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7893..7912
                     /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 cgccaacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata aatatcaata
     2401 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatcc
     2461 agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2521 taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg aagctaaaat
     2581 ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca
     2641 ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat
     2701 tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca
     2761 cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga aagacggtga
     2821 gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac
     2881 gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc
     2941 gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa
     3001 tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc
     3061 caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga
     3121 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt
     3181 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaaag
     3241 atctggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg ctgatttttg
     3301 cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag gtatgctatg
     3361 aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa ggcatatatg
     3421 atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg
     3481 ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg tttattgaaa
     3541 tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc
     3601 tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat tattgacacg
     3661 cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc
     3721 cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat gaccaccgat
     3781 atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa
     3841 aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc aggctccctt
     3901 atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt tacagtatta
     3961 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     4021 ttctcgttca gctttcttgt acaaagtggt ttacccatac gatgttccag attacgctta
     4081 gtaatgaacc ttaggagcca acagctaagc acacacttcg aatgcatctc aattagtcag
     4141 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcgtc
     4201 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     4261 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     4321 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     4381 gcttttgcaa aaagctttct agaggtacca ccggagctta tttaaataca caattaatta
     4441 acacaaatac gcgtatctgc agtcgacttt gccagaccgg agtccgaaca cagctcttcc
     4501 agcataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     4561 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     4621 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     4681 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     4741 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     4801 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     4861 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     4921 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     4981 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     5041 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcttagta
     5101 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa
     5161 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagattc cggaatttat
     5221 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaaccgg tgcagctgct ttttgcctgt
     5281 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     5341 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     5401 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     5461 agcatctaga gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5521 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5581 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5641 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     5701 ctattccaga agtagtgagg aggctttttt ggaggcctag gctagagatc ataatcagcc
     5761 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5821 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     5881 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5941 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgctagctg agcgggcttt
     6001 tttttcttag gccttcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6061 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6121 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6181 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6241 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6301 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6361 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6421 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6481 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6541 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6601 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6661 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6721 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6781 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6841 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6901 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6961 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7021 cctgactcct gcgcagtcca aaaaaaaagg ctccaaaagg agcctttaat tgtatcggtg
     7081 ggcccttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     7141 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     7201 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     7261 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     7321 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
     7381 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     7441 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     7501 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     7561 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
     7621 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     7681 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     7741 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     7801 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     7861 cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
     7921 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     7981 catcagagat tttgagacac aacgtggttt aaacaaatag tcaaaagcct ccgggcgact
     8041 agtcggggtc attagttcat agcccatata tggagttccg
//
