LOCUS       sequence-270527-        8128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral destination vector with CAMKIIA promoter, C-terminal V5 
            tag, and SV40 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8128)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8128)
  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            "Unpublished"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_8.2_M3_CAMKIIA-gateway-V5-SV40
FEATURES             Location/Qualifiers
     source          1..8128
                     /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        1078..2367
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     protein_bind    2379..2503
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2528..2533,2534..2551,2552..2558)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2612..3271
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2678..2697)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3613..3918
                     /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     3797..3816
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3959..4083)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4085..4126
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        4221..4431
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4243..4439
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4290..4425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    4554..5142
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4607..4627)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5025..5036)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5332..5512
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        5581..5791
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5650..5785
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5901..6022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5992..6011
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6074..6091)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6245..6833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6325..6344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(7085..7124)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(7134..7949)
                     /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(7134..7949)
                     /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     7857..7876
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7941..7960
                     /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 agatatccat
     1081 tatggcctta ggtcacttca tctccatggg gttcttcttc tgattttcta gaaaatgaga
     1141 tgggggtgca gagagcttcc tcagtgacct gcccagggtc acatcagaaa tgtcagagct
     1201 agaacttgaa ctcagattac taatcttaaa ttccatgcct tgggggcatg caagtacgat
     1261 atacagaagg agtgaactca ttagggcaga tgaccaatga gtttaggaaa gaagagtcca
     1321 gggcagggta catctacacc acccgcccag ccctgggtga gtccagccac gttcacctca
     1381 ttatagttgc ctctctccag tcctaccttg acgggaagca caagcagaaa ctgggacagg
     1441 agccccagga gaccaaatct tcatggtccc tctgggagga tgggtgggga gagctgtggc
     1501 agaggcctca ggaggggccc tgctgctcag tggtgacaga taggggtgag aaagcagaca
     1561 gagtcattcc gtcagcattc tgggtctgtt tggtacttct tctcacgcta aggtggcggt
     1621 gtgatatgca caatggctaa aaagcaggga gagctggaaa gaaacaagga cagagacaga
     1681 ggccaagtca accagaccaa ttcccagagg aagcaaagaa accattacag agactacaag
     1741 ggggaaggga aggagagatg aattagcttc ccctgtaaac cttagaaccc agctgttgcc
     1801 agggcaacgg ggcaatacct gtctcttcag aggagatgaa gttgccaggg taactacatc
     1861 ctgtctttct caaggaccat cccagaatgt ggcacccact agccgttacc atagcaactg
     1921 cctctttgcc ccacttaatc ccatcccgtc tgttaaaagg gccctatagt tggaggtggg
     1981 ggaggtagga agagcgatga tcacttgtgg actaagtttg ttcacatccc cttctccaac
     2041 cccctcagta catcaccctg ggagaacaag gtccacttgc ttctgggccc acacagtcct
     2101 gcagtattgt gtatataagg ccagggcaac ggaggagcag gttttgaagt gaaaggcagg
     2161 caggtgttgg ggaggcagtt accggggcaa cgggaacagg gcgtttcgga ggtggttgcc
     2221 atggggacct ggatgctgac gaaggctcgc gaggctgtga gcagccacag tgccctgctc
     2281 agaagcccca agctcgtcaa tcaagctggt tctccatttg cactcaggag cacgggcagg
     2341 cgagtggccc ctagttctgg gggcagcggg gcgcgccaac aagtttgtac aaaaaagctg
     2401 aacgagaaac gtaaaatgat ataaatatca atatattaaa ttagattttg cataaaaaac
     2461 agactacata atactgtaaa acacaacata tccagtcact atggcggccg cattaggcac
     2521 cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga gttaggatcc
     2581 gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac
     2641 cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca
     2701 atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa
     2761 aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca
     2821 tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc
     2881 ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca
     2941 cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa
     3001 cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg
     3061 ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt
     3121 tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca
     3181 ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca
     3241 gtactgcgat gagtggcagg gcggggcgta aagatctgga tccggcttac taaaagccag
     3301 ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata ctgatatgta
     3361 tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt gacagttgac
     3421 agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt ctggtaagca
     3481 caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg
     3541 aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct gacgagaaca
     3601 ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg
     3661 tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt gatccccctg
     3721 gccagtgcac gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc
     3781 ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc
     3841 ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg
     3901 atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc aggtcgacca
     3961 tagtgactgg atatgttgtg ttttacagta ttatgtagtc tgttttttat gcaaaatcta
     4021 atttaatata ttgatattta tatcatttta cgtttctcgt tcagctttct tgtacaaagt
     4081 ggttggtaag cctatcccta accctctcct cggtctcgat tctacgtagt aatgaacctt
     4141 aggagccaac agctaagcac acacttcgaa tgcatctcaa ttagtcagca accaggtgtg
     4201 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcgtctc aattagtcag
     4261 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     4321 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     4381 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     4441 agctttctag aggtaccacc ggagcttatt taaatacaca attaattaac acaaatacgc
     4501 gtatctgcag tcgactttgc cagaccggag tccgaacaca gctcttccag cataatcaac
     4561 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     4621 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     4681 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     4741 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     4801 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     4861 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     4921 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     4981 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5041 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     5101 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcttagtact ggtaccttta
     5161 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     5221 ctggaagggc taattcactc ccaacgaaga caagattccg gaatttattt gtgaaatttg
     5281 tgatgctatt gctttatttg taaaccggtg cagctgcttt ttgcctgtac tgggtctctc
     5341 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     5401 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     5461 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag catctagagt
     5521 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     5581 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     5641 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     5701 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     5761 tagtgaggag gcttttttgg aggcctaggc tagagatcat aatcagccat accacatttg
     5821 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     5881 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     5941 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     6001 ccaaactcat caatgtatct tatcatgtct gctagctgag cgggcttttt tttcttaggc
     6061 cttcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6121 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7021 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactcctgc
     7081 gcagtccaaa aaaaaaggct ccaaaaggag cctttaattg tatcggtggg cccttagaaa
     7141 aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat
     7201 ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg
     7261 gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat
     7321 ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc
     7381 ggtgagaatg gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta
     7441 cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga
     7501 gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac
     7561 cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct
     7621 aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga
     7681 gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg
     7741 accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct
     7801 ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg
     7861 cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg cggcctcgag
     7921 caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     7981 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     8041 tgagacacaa cgtggtttaa acaaatagtc aaaagcctcc gggcgactag tcggggtcat
     8101 tagttcatag cccatatatg gagttccg
//
