LOCUS       sequence-309612-        9643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector of constitutive expression of CloGFP(1-10).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9643)
  AUTHORS   Zhou S, Feng S, Brown D, Huang B
  TITLE     Improved yellow-green split fluorescent proteins for protein 
            labeling and signal amplification.
  JOURNAL   PLoS One. 2020 Nov 23;15(11):e0242592. doi: 
            10.1371/journal.pone.0242592. eCollection 2020.
  PUBMED    33227014
REFERENCE   2  (bases 1 to 9643)
  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 One.";
            date: "2020-11-23"; volume: "15(11):e0242592. doi"; pages: " 
            10.1371/journal.pone.0242592. eCollection 2020"
COMMENT     Sequence Label: pSFFV_CloGFP(1-10)
FEATURES             Location/Qualifiers
     source          1..9643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    47..172
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    669..902
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1087..1131
                     /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    1416..1533
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1686..2093
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     misc_feature    2873..3461
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2926..2946)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3344..3355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3464..3480)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3465..3481)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3571..3804
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        complement(3829..3847)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3830..3847)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4137..4156)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4370..4389)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4489..4511
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4549..4567)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4635..4739
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4740..4808,4809..5600)
                     /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(4958..4977)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5771..6359
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6260..6279
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6513..6530
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(6600..6620)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6605..6934
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6785..6920
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          8068..8133
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             8263..8283
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    8708..8842
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8745..8764)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8799..8818
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             9010..9643
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
ORIGIN
        1 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
       61 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
      121 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
      181 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
      241 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
      301 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
      361 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
      421 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
      481 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
      541 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
      601 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
      661 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
      721 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
      781 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
      841 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
      901 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
      961 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
     1021 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
     1081 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
     1141 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
     1201 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
     1261 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
     1321 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
     1381 cgacggtatc gccaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
     1441 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
     1501 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
     1561 ccagtttgga tcgataagct tgatatcgaa ttcctgcagc cccgataaaa taaaagattt
     1621 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     1681 ctgcagtaac gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca
     1741 gatcaagggc gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag
     1801 cagtttcggc cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc
     1861 gaggccaaga acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca
     1921 tcagatgttt ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac
     1981 caatcagcct gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag
     2041 ctcacaaccc ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggggggatc
     2101 tggagctctc gagaattctc acgcgtctgc aggatatcaa gcttgcggta ccgcgggccc
     2161 gggatcccat gtccaaagga gaagaactgt ttaccggtgt tgtgccaatt ttggttgaac
     2221 tcgatggtga tgtcaacgga cataagttct cagtgagagg ccaaggagaa ggtgacgcca
     2281 ccatcggaaa attgactctt aaattaatct gtactactgg taaacttcct gtaccatggc
     2341 cgactctcgt aacaacgttt gggtacggag ttgcgtgctt ttcgagatac ccagaccata
     2401 tgaaacaaca tgactttttt aagtcggcta tgcctgaagg ttacgtgcaa gaaagaacaa
     2461 tttcgttcag agatgatgga aaatataaaa ctagagcagt tgttaaattt gaaggagata
     2521 ctttggttaa ccgcattgaa ctgaaaggaa cagattttaa agaagatggt aatattcttg
     2581 gacacaaact cgaatataat tttaatagtc attacgtata catcactgct gataagcaaa
     2641 agaactgtat taaagcgaat ttcacagtac gccataatgt agaagatggc agtgttcaac
     2701 ttgccgacca ttaccaacaa aacaccccta ttggtgacgg tccggtactt cttcctgata
     2761 atcactacct cttacatcaa acaaagctga gcaaagatcc aaatgaaaaa tgagcggccg
     2821 cgactctaga gtcgacctgc aggcatgcaa gcttgatatc aagcttatcg ataatcaacc
     2881 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2941 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3001 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3061 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3121 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3181 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3241 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
     3301 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3361 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3421 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg
     3481 agggaattaa ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc
     3541 ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac
     3601 aagatctgct ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc
     3661 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     3721 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     3781 agtcagtgtg gaaaatctct agcagcatct agaattaatt ccgtgtattc tatagtgtca
     3841 cctaaatcgt atgtgtatga tacataaggt tatgtattaa ttgtagccgc gttctaacga
     3901 caatatgtac aagcctaatt gtgtagcatc tggcttactg aagcagaccc tatcatctct
     3961 ctcgtaaact gccgtcagag tcggtttggt tggacgaacc ttctgagttt ctggtaacgc
     4021 cgtcccgcac ccggaaatgg tcagcgaacc aatcagcagg gtcatcgcta gccagatcct
     4081 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     4141 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     4201 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     4261 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     4321 cctaatgcag gagtcgcata agggagagcg tcgaatggtg cactctcagt acaatctgct
     4381 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     4441 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4501 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     4561 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     4621 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4681 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4741 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4801 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4861 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4921 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4981 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5041 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5101 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5161 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5221 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5281 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5341 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5401 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5461 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5521 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5581 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5641 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5701 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5761 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5821 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5881 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5941 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6001 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6061 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6121 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6181 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6241 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6301 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6361 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6421 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6481 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     6541 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgtg
     6601 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     6661 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     6721 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     6781 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     6841 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     6901 aggaggcttt tttggaggcc taggcttttg caaaaagctt ggacacaaga caggcttgcg
     6961 agatatgttt gagaatacca ctttatcccg cgtcagggag aggcagtgcg taaaaagacg
     7021 cggactcatg tgaaatactg gtttttagtg cgccagatct ctataatctc gcgcaaccta
     7081 ttttcccctc gaacactttt taagccgtag ataaacaggc tgggacactt cacatgagcg
     7141 aaaaatacat cgtcacctgg gacatgttgc agatccatgc acgtaaactc gcaagccgac
     7201 tgatgccttc tgaacaatgg aaaggcatta ttgccgtaag ccgtggcggt ctgtaccggg
     7261 tgcgttactg gcgcgtgaac tgggtattcg tcatgtcgat accgtttgta tttccagcta
     7321 cgatcacgac aaccagcgcg agcttaaagt gctgaaacgc gcagaaggcg atggcgaagg
     7381 cttcatcgtt attgatgacc tggtggatac cggtggtact gcggttgcga ttcgtgaaat
     7441 gtatccaaaa gcgcactttg tcaccatctt cgcaaaaccg gctggtcgtc cgctggttga
     7501 tgactatgtt gttgatatcc cgcaagatac ctggattgaa cagccgtggg atatgggcgt
     7561 cgtattcgtc ccgccaatct ccggtcgcta atcttttcaa cgcctggcac tgccgggcgt
     7621 tgttcttttt aacttcaggc gggttacaat agtttccagt aagtattctg gaggctgcat
     7681 ccatgacaca ggcaaacctg agcgaaaccc tgttcaaacc ccgctttaaa catcctgaaa
     7741 cctcgacgct agtccgccgc tttaatcacg gcgcacaacc gcctgtgcag tcggcccttg
     7801 atggtaaaac catccctcac tggtatcgca tgattaaccg tctgatgtgg atctggcgcg
     7861 gcattgaccc acgcgaaatc ctcgacgtcc aggcacgtat tgtgatgagc gatgccgaac
     7921 gtaccgacga tgatttatac gatacggtga ttggctaccg tggcggcaac tggatttatg
     7981 agtgggcccc ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa
     8041 ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt
     8101 aaactactga ttctaattgt ttgtgtattt tagattccaa cctatggaac tgatgaatgg
     8161 gagcagtggt ggaatgcctt taatgaggaa aacctgtttt gctcagaaga aatgccatct
     8221 agtgatgatg aggctactgc tgactctcaa cattctactc ctccaaaaaa gaagagaaag
     8281 gtagaagacc ccaaggactt tccttcagaa ttgctaagtt ttttgagtca tgctgtgttt
     8341 agtaatagaa ctcttgcttg ctttgctatt tacaccacaa aggaaaaagc tgcactgcta
     8401 tacaagaaaa ttatggaaaa atattctgta acctttataa gtaggcataa cagttataat
     8461 cataacatac tgttttttct tactccacac aggcatagag tgtctgctat taataactat
     8521 gctcaaaaat tgtgtacctt tagcttttta atttgtaaag gggttaataa ggaatatttg
     8581 atgtatagtg ccttgactag agatcataat cagccatacc acatttgtag aggttttact
     8641 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     8701 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8761 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8821 tgtatcttat catgtctgga tcaactggat aactcaagct aaccaaaatc atcccaaact
     8881 tcccacccca taccctatta ccactgccaa ttacctgtgg tttcatttac tctaaacctg
     8941 tgattcctct gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa
     9001 cttagtagtt ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga
     9061 tctaccacac acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca
     9121 gatatccact gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag
     9181 aagaggccaa taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg
     9241 atgacccgga gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg
     9301 tggcccgaga gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg
     9361 gactttccgc tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag
     9421 ccctcagatc ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac
     9481 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     9541 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     9601 agatccctca gaccctttta gtcagtgtgg aaaatctcta gca
//
