LOCUS       sequence-270524-        7015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral destination vector with hcSyn promoter and C-terminal V5 
            tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7015)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7015)
  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            "Unpublished"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_7.1_M2_hcSyn-gateway-V5
FEATURES             Location/Qualifiers
     source          1..7015
                     /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        1100..1547
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    1557..1681
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1706..1711,1712..1729,1730..1736)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1790..2449
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1856..1875)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2791..3096
                     /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     2975..2994
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3137..3261)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3263..3304
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     misc_feature    3441..4029
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3494..3514)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3912..3923)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4219..4399
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        4468..4678
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4537..4672
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4788..4909
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4879..4898
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4961..4978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5132..5720)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5212..5231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5972..6011)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(6021..6836)
                     /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(6021..6836)
                     /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     6744..6763
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6828..6847
                     /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 agatatctct
     1081 gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg
     1141 gtgcctacct gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa
     1201 ttgcgcatcc cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac
     1261 tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc
     1321 gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg
     1381 gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg
     1441 agataggggg gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc
     1501 agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag agcgcagggc gcgccaacaa
     1561 gtttgtacaa aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     1621 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat
     1681 ggcggccgca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
     1741 gattttgagt taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa
     1801 aatcactgga tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc
     1861 atttcagtca gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt
     1921 tttaaagacc gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc
     1981 ccgcctgatg aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat
     2041 atgggatagt gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc
     2101 gctctggagt gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt
     2161 ggcgtgttac ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt
     2221 cgtctcagcc aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga
     2281 caacttcttc gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct
     2341 gatgccgctg gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat
     2401 gcttaatgaa ttacaacagt actgcgatga gtggcagggc ggggcgtaaa gatctggatc
     2461 cggcttacta aaagccagat aacagtatgc gtatttgcgc gctgattttt gcggtataag
     2521 aatatatact gatatgtata cccgaagtat gtcaaaaaga ggtatgctat gaagcagcgt
     2581 attacagtga cagttgacag cgacagctat cagttgctca aggcatatat gatgtcaata
     2641 tctccggtct ggtaagcaca accatgcaga atgaagcccg tcgtctgcgt gccgaacgct
     2701 ggaaagcgga aaatcaggaa gggatggctg aggtcgcccg gtttattgaa atgaacggct
     2761 cttttgctga cgagaacagg ggctggtgaa atgcagttta aggtttacac ctataaaaga
     2821 gagagccgtt atcgtctgtt tgtggatgta cagagtgata ttattgacac gcccgggcga
     2881 cggatggtga tccccctggc cagtgcacgt ctgctgtcag ataaagtctc ccgtgaactt
     2941 tacccggtgg tgcatatcgg ggatgaaagc tggcgcatga tgaccaccga tatggccagt
     3001 gtgccggtct ccgttatcgg ggaagaagtg gctgatctca gccaccgcga aaatgacatc
     3061 aaaaacgcca ttaacctgat gttctgggga atataaatgt caggctccct tatacacagc
     3121 cagtctgcag gtcgaccata gtgactggat atgttgtgtt ttacagtatt atgtagtctg
     3181 ttttttatgc aaaatctaat ttaatatatt gatatttata tcattttacg tttctcgttc
     3241 agctttcttg tacaaagtgg ttggtaagcc tatccctaac cctctcctcg gtctcgattc
     3301 tacgtagtaa tgaaccttag gagccaacag ctaagcacac acttcgaaag atatatttaa
     3361 atacacaatt aattaacaca aatacgcgta tctgcagtcg actttgccag accggagtcc
     3421 gaacacagct cttccagcat aatcaacctc tggattacaa aatttgtgaa agattgactg
     3481 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     3541 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     3601 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     3661 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     3721 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     3781 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaaatcat
     3841 cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     3901 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     3961 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4021 cctccccgct tagtactggt acctttaaga ccaatgactt acaaggcagc tgtagatctt
     4081 agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa
     4141 gattccggaa tttatttgtg aaatttgtga tgctattgct ttatttgtaa accggtgcag
     4201 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     4261 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     4321 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     4381 gtgtggaaaa tctctagcat ctagagtatg caaagcatgc atctcaatta gtcagcaacc
     4441 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4501 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     4561 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     4621 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggctag
     4681 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4741 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4801 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4861 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgct
     4921 agctgagcgg gctttttttt cttaggcctt cttccgcttc ctcgctcact gactcgctgc
     4981 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     5041 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     5101 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5161 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5221 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5281 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5341 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5401 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5461 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5521 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5581 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5641 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5701 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5761 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5821 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5881 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5941 gttcatccat agttgcctga ctcctgcgca gtccaaaaaa aaaggctcca aaaggagcct
     6001 ttaattgtat cggtgggccc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     6061 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     6121 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     6181 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     6241 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     6301 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     6361 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     6421 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     6481 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     6541 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     6601 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     6661 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     6721 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     6781 atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcataaca
     6841 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     6901 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggtttaaaca aatagtcaaa
     6961 agcctccggg cgactagtcg gggtcattag ttcatagccc atatatggag ttccg
//
