LOCUS       sequence-270528-        7837 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral destination vector with CAMKIIA promoter and C-terminal 
            V5 tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7837)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7837)
  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            "Unpublished"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_8.1_ M2_CAMKIIA-gateway-V5
FEATURES             Location/Qualifiers
     source          1..7837
                     /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"
     misc_feature    4263..4851
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4316..4336)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4734..4745)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5041..5221
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        5290..5500
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5359..5494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5610..5731
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5701..5720
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5783..5800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5954..6542)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6034..6053)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6794..6833)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(6843..7658)
                     /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(6843..7658)
                     /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     7566..7585
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7650..7669
                     /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 agatatattt aaatacacaa ttaattaaca
     4201 caaatacgcg tatctgcagt cgactttgcc agaccggagt ccgaacacag ctcttccagc
     4261 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4321 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4381 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
     4441 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
     4501 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     4561 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     4621 tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc
     4681 tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc
     4741 tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc
     4801 ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg cttagtactg
     4861 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa
     4921 aaggggggac tggaagggct aattcactcc caacgaagac aagattccgg aatttatttg
     4981 tgaaatttgt gatgctattg ctttatttgt aaaccggtgc agctgctttt tgcctgtact
     5041 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
     5101 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
     5161 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
     5221 atctagagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5281 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5341 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     5401 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     5461 ttccagaagt agtgaggagg cttttttgga ggcctaggct agagatcata atcagccata
     5521 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga
     5581 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca
     5641 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5701 gtggtttgtc caaactcatc aatgtatctt atcatgtctg ctagctgagc gggctttttt
     5761 ttcttaggcc ttcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5821 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5881 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5941 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6001 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6061 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6121 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6181 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6241 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6301 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6361 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6421 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6481 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6541 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6601 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6661 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6721 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6781 gactcctgcg cagtccaaaa aaaaaggctc caaaaggagc ctttaattgt atcggtgggc
     6841 ccttagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     6901 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     6961 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     7021 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     7081 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     7141 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     7201 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     7261 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     7321 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     7381 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     7441 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     7501 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     7561 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     7621 ggcctcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
     7681 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
     7741 cagagatttt gagacacaac gtggtttaaa caaatagtca aaagcctccg ggcgactagt
     7801 cggggtcatt agttcatagc ccatatatgg agttccg
//
