LOCUS       sequence-228426-        6740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral transfer vector containing a multible cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  AUTHORS   Wang R, Pscheid R, Ghumra A, Kan LYL, Cochrane S, Fairclough L, 
            Alcocer MJC
  TITLE     Towards a surrogate system to express human lipid binding TCRs.
  JOURNAL   Biotechnol Lett. 2019 Oct;41(10):1095-1104. doi: 
            10.1007/s10529-019-02713-2. Epub 2019 Jul 26.
  PUBMED    31346817
REFERENCE   2  (bases 1 to 6740)
  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"; doi: 
            "10.1007/s10529-019-02713-2"; journalName: "Biotechnol Lett"; date: 
            "2019-10"; volume: "41"; issue: "10"; pages: "1095-1104"
COMMENT     Sequence Label: pMJA284= pSin-Pur-MCS-Puro
FEATURES             Location/Qualifiers
     source          1..6740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             118..751
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    798..923
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1416..1649
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1834..1878
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2165..2282
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2430..2641
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    2857..3430
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3024..3041)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3251..3270
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3463..4062
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3463..3482)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3959..3979
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4265..4498
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4649..4666)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4655..4879
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      4922..4931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        4975..5079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5080..5148,5149..5940)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5298..5317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6111..6699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6600..6619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggaccgcc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
       61 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagttgg
      121 aagggctaat tcactcccaa agaagacaag atatccttga tctgtggatc taccacacac
      181 aaggctactt ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga
      241 cctttggatg gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata
      301 aaggagagaa caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga
      361 gagaagtgtt agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc
      421 tgcatccgga gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg
      481 gggactttcc agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct
      541 gcatataagc agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc
      601 tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga
      661 gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga
      721 cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga
      781 aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
      841 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
      901 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
      961 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     1021 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     1081 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     1141 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     1201 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     1261 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     1321 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     1381 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     1441 ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa
     1501 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     1561 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     1621 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     1681 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     1741 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     1801 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     1861 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     1921 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     1981 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     2041 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     2101 agatccattc gattagtgaa cggatctcga cggtatcgcc acaaatggca gtattcatcc
     2161 acaattttaa aagaaagggg gggattgggg ggtacagtgc aggggaaaga atagtagaca
     2221 taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa attcaaaatt
     2281 ttcgggttta ttacagggac agcagagatc cactttggat cgataagctt tgcaaagatg
     2341 gataaagttt taaacagaga ggaatctttg cagctaatgg accttctagg tcttgaaagg
     2401 agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     2461 gaagttgggg ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa
     2521 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     2581 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     2641 ggtaagtgcc gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt
     2701 gccttgaatt acttccacct ggctgcagta atttaaaact agtaatttcc taggtagttt
     2761 aaacaagaaa cttttgaatt cgaagcttga gctcgagatc tgagtccggt agcgctagtt
     2821 cgaaggatcc gcatgcatct agggcggcca attccgcccc tctccccccc ccccccctaa
     2881 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     2941 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     3001 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     3061 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     3121 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     3181 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     3241 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     3301 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     3361 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     3421 acgatgataa gcttgccaca acccacaagg agacgacctt ccatgaccga gtacaagccc
     3481 acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg tacgcaccct cgccgccgcg
     3541 ttcgccgact accccgccac gcgccacacc gtcgacccgg accgccacat cgagcgggtc
     3601 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     3661 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     3721 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     3781 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     3841 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     3901 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc
     3961 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     4021 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gataataggc ggccgctcga
     4081 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     4141 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     4201 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     4261 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     4321 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     4381 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     4441 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     4501 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     4561 agtgagaggc cttgacatta taatagattt agcaggaatt gaactaggag tggagcacac
     4621 aggcaaagtt ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     4681 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     4741 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     4801 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     4861 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctgggggcg cgcccctcga
     4921 ggccgccatg gtcatagctg tttgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4981 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcac gagacaataa
     5041 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5101 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5161 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5221 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5281 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5341 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5401 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5461 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5521 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5581 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5641 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5701 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5761 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5821 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5881 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5941 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6001 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6061 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6121 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6181 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6241 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6301 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6361 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6421 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6481 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6541 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6601 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6661 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6721 ttacggttct ggccttttgc
//
