LOCUS       sequence-256447-        7592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral expression vector with CAMKIIA promoter, EGFP gene, 
            C-terminal V5 tag, SV40 promoter, and Zeo Resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7592)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7592)
  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            "Unpublished"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_8.4_Ex_CAMKIIA-EGFP-V5-SV40-Zeo
FEATURES             Location/Qualifiers
     source          1..7592
                     /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..2403
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      2424..2433
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2430..2432,2433..2435,2436..3146)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2475..2496)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2736..2755)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3083..3104
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3148..3172)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3174..3215
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     promoter        3310..3520
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        3332..3528
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3379..3514
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3565..3574
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3571..3945
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    4018..4606
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4071..4091)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4489..4500)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4796..4976
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        5045..5255
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5114..5249
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5365..5486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     5456..5475
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5538..5555)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5709..6297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5789..5808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6549..6588)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(6598..7413)
                     /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(6598..7413)
                     /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     7321..7340
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     7405..7424
                     /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 aaaaaagcag
     2401 gctaagctgg ctagttaagc ttcgccacca tggtgagcaa gggcgaggag ctgttcaccg
     2461 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2521 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2581 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     2641 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     2701 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     2761 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     2821 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     2881 tctatatcat ggccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca
     2941 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3001 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3061 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3121 ctctcggcat ggacgagctg tacaagcacc cagctttctt gtacaaagtg gttggtaagc
     3181 ctatccctaa ccctctcctc ggtctcgatt ctacgtagta atgaacctta ggagccaaca
     3241 gctaagcaca cacttcgaat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3301 ggctccccag caggcagaag tatgcaaagc atgcgtctca attagtcagc aaccatagtc
     3361 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     3481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctttctaga
     3541 ggtaccaccg gagcttattt aaatgccacc atggccaagt tgaccagtgc cgttccggtg
     3601 ctcaccgcgc gcgacgtcgc cggagcggtc gagttctgga ccgaccggct cgggttctcc
     3661 cgggacttcg tggaggacga cttcgccggt gtggtccggg acgacgtgac cctgttcatc
     3721 agcgcggtcc aggaccaggt ggtgccggac aacaccctgg cctgggtgtg ggtgcgcggc
     3781 ctggacgagc tgtacgccga gtggtcggag gtcgtgtcca cgaacttccg ggacgcctcc
     3841 gggccggcca tgaccgagat cggcgagcag ccgtgggggc gggagttcgc cctgcgcgac
     3901 ccggccggca actgcgtgca cttcgtggcc gaggagcagg actaattaat taacacaaat
     3961 acgcgtatct gcagtcgact ttgccagacc ggagtccgaa cacagctctt ccagcataat
     4021 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4081 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4141 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4201 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4261 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4321 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4381 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4441 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4501 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4561 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcttag tactggtacc
     4621 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     4681 gggactggaa gggctaattc actcccaacg aagacaagat tccggaattt atttgtgaaa
     4741 tttgtgatgc tattgcttta tttgtaaacc ggtgcagctg ctttttgcct gtactgggtc
     4801 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     4861 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     4921 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcatcta
     4981 gagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     5041 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     5101 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     5161 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     5221 gaagtagtga ggaggctttt ttggaggcct aggctagaga tcataatcag ccataccaca
     5281 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     5341 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     5401 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5461 ttgtccaaac tcatcaatgt atcttatcat gtctgctagc tgagcgggct tttttttctt
     5521 aggccttctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5581 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5641 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5701 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5761 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5821 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5881 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5941 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6001 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6061 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6121 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6181 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6241 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6301 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6361 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6421 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6481 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6541 ctgcgcagtc caaaaaaaaa ggctccaaaa ggagccttta attgtatcgg tgggccctta
     6601 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     6661 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     6721 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     6781 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     6841 atccggtgag aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc
     6901 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     6961 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     7021 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     7081 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     7141 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     7201 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     7261 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     7321 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     7381 cgagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     7441 agcagacagt tttattgttc atgatgatat atttttatct tgtgcaatgt aacatcagag
     7501 attttgagac acaacgtggt ttaaacaaat agtcaaaagc ctccgggcga ctagtcgggg
     7561 tcattagttc atagcccata tatggagttc cg
//
