LOCUS       sequence-335072-        8222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green lentiviral vector plasmid for pseudovirus experiments.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8222)
  AUTHORS   Shukla N, Roelle SM, Suzart VG, Bruchez AM, Matreyek KA
  TITLE     Mutants of human ACE2 differentially promote SARS-CoV and SARS-CoV-2
            spike mediated infection.
  JOURNAL   PLoS Pathog. 2021 Jul 16;17(7):e1009715. doi: 
            10.1371/journal.ppat.1009715. eCollection 2021 Jul.
  PUBMED    34270613
REFERENCE   2  (bases 1 to 8222)
  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: "PLoS 
            Pathog."; date: "2021-07-16"; volume: "17(7):e1009715. doi"; pages: 
            " 10.1371/journal.ppat.1009715. eCollection 2021 Jul"
COMMENT     Sequence Label: pLenti_CMV-EGFP-2A-mNeonGreen
FEATURES             Location/Qualifiers
     source          1..8222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..24,25..31)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    39..55
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     44..66
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     63..79
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     63..79
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     98..118
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        100..118
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        146..372
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             373..553
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    600..725
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1218..1451
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1636..1680
                     /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    1947..2063
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2086..2389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2390..2593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2544..2564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2590..2614
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2625..2634
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2676..2697)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2937..2956)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3284..3305
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3339..3395
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     regulatory      3648..3657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(4106..4130)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4161..4749
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4214..4234)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4632..4643)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4886..5119
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5191..5312
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5228..5247)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5282..5301
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5352..5487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5507..5526)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5508..5526)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5536..5553)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5536..5552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5545..5567)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5694..6149
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5781..5800)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5991..6012
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6175..6279
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6280..6348,6349..7140)
                     /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(6498..6517)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7311..7899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7800..7819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8053..8070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8187..8208
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
       61 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga
      121 acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta gtcttgcaac
      181 atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg tgcatgccga
      241 ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca
      301 tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa gtgcctagct
      361 cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
      421 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
      481 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
      541 aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc
      601 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
      661 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
      721 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
      781 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
      841 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
      901 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
      961 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     1021 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     1081 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     1141 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     1201 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     1261 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     1321 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     1381 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     1441 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     1501 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     1561 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     1621 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     1681 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     1741 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     1801 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     1861 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     1921 aacggatctc gacggtatcg gttaactttt aaaagaaaag gggggattgg ggggtacagt
     1981 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     2041 caaattacaa aattcaaaat tttatcgata agcttgggag ttccgcgtta cataacttac
     2101 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     2161 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     2221 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     2281 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     2341 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     2401 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     2461 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     2521 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     2581 tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccacc atggtgtcta
     2641 aaggagaaga attatttacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2701 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2761 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2821 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2881 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2941 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3001 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3061 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     3121 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     3181 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     3241 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3301 tcgtgaccgc cgccgggatc actctcggag gtagcggagc tactaacttc agcctgctga
     3361 agcaggctgg agacgtggag gagaaccctg gacctgtgtc caagggcgag gaggacaaca
     3421 tggcctccct gcccgctacc cacgagctgc acattttcgg ctccattaac ggcgtggact
     3481 tcgacatggt gggccagggc acaggcaacc caaacgacgg ctacgaggag ctgaacctga
     3541 agtctacaaa gggcgacctg cagttctctc catggatcct ggtgccccac atcggctacg
     3601 gcttccacca gtacctcccc taccccgacg gcatgtcccc tttccaggcc gccatggtgg
     3661 acggcagcgg ctaccaggtg cacaggacca tgcagttcga ggacggcgct agcctgacag
     3721 tgaactaccg ctacacctac gagggcagcc acattaaggg cgaggctcag gtgaagggca
     3781 caggcttccc tgccgacggc cctgtgatga caaactctct gacagccgct gactggtgca
     3841 gatctaagaa gacctaccca aacgacaaga ccatcattag caccttcaag tggtcttaca
     3901 ccaccggcaa cggcaagcgg tacagaagca cagccaggac aacatacaca ttcgctaagc
     3961 ccatggctgc taactacctg aagaaccagc ccatgtacgt gttcaggaag acagagctga
     4021 agcacagcaa gacagagctc aacttcaagg agtggcagaa ggctttcacc gacgtgatgg
     4081 gcatggacga gctgtacaag taatcaccca gctttcttgt acaaagtggt tgatatccag
     4141 cacagtggcg gccgctcgac aatcaacctc tggattacaa aatttgtgaa agattgactg
     4201 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atgcctttgt
     4261 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
     4321 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
     4381 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
     4441 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
     4501 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
     4561 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
     4621 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
     4681 tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc ctttgggccg
     4741 cctccccgcc tggaattctg cagatatccg gttagtaatg agtttggaat taattctgtg
     4801 cacaattcga gctcggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
     4861 cactttttaa aagaaaaggg gggactggaa gggctagctc actcccaacg aagacaagat
     4921 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     4981 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     5041 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     5101 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
     5161 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
     5221 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     5281 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
     5341 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5401 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5461 agtagtgagg aggctttttt ggaggcctag ggacgtaccc aattcgccct atagtgagtc
     5521 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     5581 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     5641 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc
     5701 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     5761 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     5821 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     5881 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     5941 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6001 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6061 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     6121 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     6181 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6241 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6301 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6361 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6421 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6481 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6541 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6601 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6661 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6721 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6781 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6841 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6901 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6961 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     7021 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     7081 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7141 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7201 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7261 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7321 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7381 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7441 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7501 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7561 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7621 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7681 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7741 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7801 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7861 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7921 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7981 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     8041 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     8101 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     8161 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     8221 gc
//
