LOCUS       sequence-256453-        6536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral destination vector with C-terminal HA tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6536)
  TITLE     Barrett lab unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6536)
  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            "Unpublished"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_6.3_M1-gateway-HA
FEATURES             Location/Qualifiers
     source          1..6536
                     /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"
     protein_bind    1093..1217
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1242..1247,1248..1265,1266..1272)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1326..1985
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1392..1411)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2327..2632
                     /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     2511..2530
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2673..2797)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2799..2825
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2962..3550
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3015..3035)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3433..3444)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3740..3920
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        3989..4199
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4058..4193
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4309..4430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     4400..4419
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4482..4499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4653..5241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4733..4752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(5493..5532)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(5542..6357)
                     /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(5542..6357)
                     /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     6265..6284
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6349..6368
                     /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 agatatcccg
     1081 caaggcgcgc caacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     1141 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     1201 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     1261 gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga gctaaggaag
     1321 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     1381 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     1441 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     1501 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     1561 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     1621 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     1681 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     1741 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     1801 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     1861 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt tgtgatggct
     1921 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     1981 cgtaaagatc tggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     2041 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     2101 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     2161 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     2221 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     2281 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     2341 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     2401 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     2461 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     2521 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     2581 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     2641 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     2701 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     2761 tttacgtttc tcgttcagct ttcttgtaca aagtggttta cccatacgat gttccagatt
     2821 acgcttagta atgaacctta ggagccaaca gctaagcaca cacttcgaaa gatatattta
     2881 aatacacaat taattaacac aaatacgcgt atctgcagtc gactttgcca gaccggagtc
     2941 cgaacacagc tcttccagca taatcaacct ctggattaca aaatttgtga aagattgact
     3001 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3061 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3121 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3181 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3241 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3301 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3361 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3421 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3481 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3541 gcctccccgc ttagtactgg tacctttaag accaatgact tacaaggcag ctgtagatct
     3601 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca
     3661 agattccgga atttatttgt gaaatttgtg atgctattgc tttatttgta aaccggtgca
     3721 gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     3781 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     3841 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     3901 agtgtggaaa atctctagca tctagagtat gcaaagcatg catctcaatt agtcagcaac
     3961 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     4021 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4081 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4141 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggcta
     4201 gagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     4261 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     4321 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4381 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgc
     4441 tagctgagcg ggcttttttt tcttaggcct tcttccgctt cctcgctcac tgactcgctg
     4501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     4861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5461 cgttcatcca tagttgcctg actcctgcgc agtccaaaaa aaaaggctcc aaaaggagcc
     5521 tttaattgta tcggtgggcc cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     5581 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     5641 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     5701 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     5761 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt
     5821 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     5881 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     5941 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     6001 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     6061 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     6121 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     6181 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     6241 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     6301 catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat ggctcataac
     6361 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
     6421 atcttgtgca atgtaacatc agagattttg agacacaacg tggtttaaac aaatagtcaa
     6481 aagcctccgg gcgactagtc ggggtcatta gttcatagcc catatatgga gttccg
//
