LOCUS       sequence-377150-        9020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector bearing a hybrid MSCV/HIV-1 3'-LTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9020)
  AUTHORS   Zhang XY, La Russa VF, Reiser J
  TITLE     Transduction of bone-marrow-derived mesenchymal stem cells by using 
            lentivirus vectors pseudotyped with modified RD114 envelope 
            glycoproteins.
  JOURNAL   J Virol. 2004 Feb . 78(3):1219-29.
  PUBMED    14722277
REFERENCE   2  (bases 1 to 9020)
  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: "J Virol. 
            2004 Feb . 78(3):1219-29."
COMMENT     Sequence Label: pNL-EGFP(MSCV)
FEATURES             Location/Qualifiers
     source          1..9020
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..31
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(250..354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     422..440
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(478..500)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     600..619
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     LTR             2312..2569
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      3196..3331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     misc_feature    3609..3842
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             4027..4071
                     /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"
     regulatory      4348..4357
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4354..4356,4357..4359,4360..5070)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4354..4356,4357..4359,4360..5070)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4399..4420)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4660..4679)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5007..5028
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5659..5839
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(7479..7496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7650..8238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7730..7749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8409..9020,1..180,181..249))
                     /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"
ORIGIN
        1 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
       61 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      121 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      181 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      241 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      301 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
      361 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
      421 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
      481 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
      541 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
      601 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
      661 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
      721 agggcgatcg gtgcgggcct cttcgctatt acgccagggg aggcagagat tgcagtaagc
      781 tgagatcgca gcactgcact ccagcctggg cgacagagta agactctgtc tcaaaaataa
      841 aataaataaa tcaatcagat attccaatct tttcctttat ttatttattt attttctatt
      901 ttggaaacac agtccttcct tattccagaa ttacacatat attctatttt tctttatatg
      961 ctccagtttt ttttagacct tcacctgaaa tgtgtgtata caaaatctag gccagtccag
     1021 cagagcctaa aggtaaaaaa taaaataata aaaaataaat aaaatctagc tcactccttc
     1081 acatcaaaat ggagatacag ctgttagcat taaataccaa ataacccatc ttgtcctcaa
     1141 taattttaag cgcctctctc caccacatct aactcctgtc aaaggcatgt gccccttccg
     1201 ggcgctctgc tgtgctgcca accaactggc atgtggactc tgcagggtcc ctaactgcca
     1261 agccccacag tgtgccctga ggctgcccct tccttctagc ggctgccccc actcggcttt
     1321 gctttcccta gtttcagtta cttgcgttca gccaaggtct gaaactaggt gcgcacagag
     1381 cggtaagact gcgagagaaa gagaccagct ttacaggggg tttatcacag tgcaccctga
     1441 cagtcgtcag cctcacaggg ggtttatcac attgcaccct gacagtcgtc agcctcacag
     1501 ggggtttatc acagtgcacc cttacaatca ttccatttga ttcacaattt ttttagtctc
     1561 tactgtgcct aacttgtaag ttaaatttga tcagaggtgt gttcccagag gggaaaacag
     1621 tatatacagg gttcagtact atcgcatttc aggcctccac ctgggtcttg gaatgtgtcc
     1681 cccgaggggt gatgactacc tcagttggat ctccacaggt cacagtgaca caagataacc
     1741 aagacacctc ccaaggctac cacaatgggc cgccctccac gtgcacatgg ccggaggaac
     1801 tgccatgtcg gaggtgcaag cacacctgcg catcagagtc cttggtgtgg agggagggac
     1861 cagcgcagct tccagccatc cacctgatga acagaaccta gggaaagccc cagttctact
     1921 tacaccagga aaggctggaa gggctaattt ggtcccaaaa aagacaagag atccttgatc
     1981 tgtggatcta ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag
     2041 ggatcagata tccactgacc tttggatggt gcttcaagtt agtaccagtt gaaccagagc
     2101 aagtagaaga ggccaatgaa ggagagaaca acagcttgtt acaccctatg agccagcatg
     2161 ggatggagga cccggaggga gaagtattag tgtggaagtt tgacagcctc ctagcatttc
     2221 gtcacatggc ccgagagctg catccggagt actacaaaga ctgctgacat cgagctttct
     2281 acaagggact ttccgctggg gactttccag ggaggtgtgg cctgggcggg actggggagt
     2341 ggcgagccct cagatgctac atataagcag ctgctttttg cctgtactgg gtctctctgg
     2401 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     2461 caataaagct tgccttgagt gctcaaagta gtgtgtgccc gtctgttgtg tgactctggt
     2521 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga
     2581 acagggactt gaaagcgaaa gtaaagccag aggagatctc tcgacgcagg actcggcttg
     2641 ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga
     2701 ctagcggagg ctagaaggag agagatgggt gcgagagcgt cggtattaag cgggggagaa
     2761 ttagataaat gggaaaaaat tcggttaagg ccagggggaa agaaacaata taaactaaaa
     2821 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg ccttttagag
     2881 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     2941 gaagaactta gatcattata taatacaata gcagtcctct attgtgtgca tcaaaggata
     3001 gatgtaaaag acaccaagga agccttagat aagatagagg aagagcaaaa caaaagtaag
     3061 aaaaaggcac agcaagcagc agctgacaca ggaaacaaca gccaggtcag ccaaaattac
     3121 cctatagtgc agaacctcca ggggcaaatg gtacatcagg ccatatcacc tagaacttta
     3181 aatatgactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3241 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3301 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaatgcat aaaagaaaag
     3361 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     3421 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     3481 acagcagaga tccacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
     3541 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
     3601 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
     3661 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
     3721 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
     3781 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
     3841 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
     3901 agttggagta ataaatctct ggaacagatt tggaataaca tgacctggat ggagtgggac
     3961 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     4021 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     4081 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     4141 gtaggtttaa gaatagtttt tgctgtactt tctgtagtga atagagttag gcagggatat
     4201 tcaccattat cgtttcagac ccacctccca atcccgaggg gacccgacag gcccgaagga
     4261 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     4321 agttaactag atctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc
     4381 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     4441 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
     4501 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     4561 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     4621 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     4681 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     4741 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     4801 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     4861 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     4921 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     4981 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     5041 atcactctcg gcatggacga gctgtacaag tccggactca gatccgacct cgagacctag
     5101 aaaaacatgg agcaatcaca agtagcaata cagcagctaa caatgctgct tgtgcctggc
     5161 tagaagcaca agaggaggaa gaggtgggtt ttccagtcac acctcaggta cctttaagac
     5221 caatgactta caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg
     5281 aagggctaat tcactcccaa agaagacaag atatccttga tctgtgggta acgccatttt
     5341 gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt taggaacaga
     5401 gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
     5461 gggccaagaa cagatggtcc ccagatgcgg tcccgccctc agcagtttct agagaaccat
     5521 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
     5581 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
     5641 ccacattttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     5701 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5761 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5821 gtgtggaaaa tctctagcac ccaggaggta gaggttgcag tgagccaaga tcgcgccact
     5881 gcattccagc ctgggcaaga aaacaagact gtctaaaata ataataataa gttaagggta
     5941 ttaaatatat ttatacatgg aggtcataaa aatatatata tttgggctgg gcgcagtggc
     6001 tcacacctgc gcccggccct ttgggaggcc gaggcaggtg gatcacctga gtttgggagt
     6061 tccagaccag cctgaccaac atggagaaac cccttctctg tgtattttta gtagatttta
     6121 ttttatgtgt attttattca caggtatttc tggaaaactg aaactgtttt tcctctactc
     6181 tgataccaca agaatcatca gcacagagga agacttctgt gatcaaatgt ggtgggagag
     6241 ggaggttttc accagcacat gagcagtcag ttctgccgca gactcggcgg gtgtccttcg
     6301 gttcagttcc aacaccgcct gcctggagag aggtcagacc acagggtgag ggctcagtcc
     6361 ccaagacata aacacccaag acataaacac ccaacaggtc caccccgcct gctgcccagg
     6421 cagagccgat tcaccaagac gggaattagg atagagaaag agtaagtcac acagagccgg
     6481 ctgtgcggga gaacggagtt ctattatgac tcaaatcagt ctccccaagc attcggggat
     6541 cagagttttt aaggataact tagtgtgtag ggggccagtg agttggagat gaaagcgtag
     6601 ggagtcgaag gtgtcctttt gcgccgagtc agttcctggg tgggggccac aagatcggat
     6661 gagccagttt atcaatccgg gggtgccagc tgatccatgg agtgcagggt ctgcaaaata
     6721 tctcaagcac tgattgatct taggttttac aatagtgatg ttaccccagg aacaatttgg
     6781 ggaaggtcag aatcttgtag cctgtagctg catgactcct aaaccataat ttcttttttg
     6841 tttttttttt tttatttttg agacagggtc tcactctgtc acctaggctg gagtgcagtg
     6901 gtgcaatcac agctcactgc agcctcaacg tcgtaagctc aagcgatcct cccacctcag
     6961 cctgcctggt agctgagact acaagcgacg ccccagttaa tttttgtatt tttggtagag
     7021 gcagcgtttt gccgtgtggc cctggctggt ctcgaactcc tgggctcaag tgatccagcc
     7081 tcagcctccc aaagtgctgg gacaaccggg gccagtcact gcacctggcc ctaaaccata
     7141 atttctaatc ttttggctaa tttgttagtc ctacaaaggc agtctagtcc ccaggcaaaa
     7201 agggggtttg tttcgggaaa gggctgttac tgtctttgtt tcaaactata aactaagttc
     7261 ctcctaaact tagttcggcc tacacccagg aatgaacaag gagagcttgg aggttagaag
     7321 cacgatggaa ttggttaggt cagatctctt tcactgtctg agttataatt ttgcaatggt
     7381 ggttcaaaga ctgcccgctt ctgacaccag tcgctgcatt aatgaatcgg ccaacgcgcg
     7441 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     7501 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     7561 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     7621 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     7681 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     7741 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     7801 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     7861 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     7921 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     7981 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     8041 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     8101 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     8161 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     8221 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     8281 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     8341 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     8401 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     8461 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     8521 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     8581 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     8641 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     8701 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     8761 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     8821 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     8881 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     8941 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     9001 ccgagttgct cttgcccggc
//
