LOCUS       sequence-232358-        9444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express gag-gfp fusion driven by EIF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9444)
  AUTHORS   Yurkovetskiy L, Guney MH, Kim K, Goh SL, McCauley S, Dauphin A, 
            Diehl WE, Luban J
  TITLE     Primate immunodeficiency virus proteins Vpx and Vpr counteract 
            transcriptional repression of proviruses by the HUSH complex.
  JOURNAL   Nat Microbiol. 2018 Dec;3(12):1354-1361. doi: 
            10.1038/s41564-018-0256-x. Epub 2018 Oct 8.
  PUBMED    30297740
REFERENCE   2  (bases 1 to 9444)
  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: 
            "10.1038/s41564-018-0256-x"; journalName: "Nat Microbiol"; date: 
            "2018-12"; volume: "3"; issue: "12"; pages: "1354-1361"
COMMENT     Sequence Label: pecALPS gag-gfp Blasti
FEATURES             Location/Qualifiers
     source          1..9444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(64..84)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        69..398
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      249..384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     311..330
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        410..789
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        791..989
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     944..964
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1007..1187
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1234..1359
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1856..2089
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2273..2317
                     /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    2615..2732
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2789..3000
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      3007..3016
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3011..3020
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4531..4533,4534..4536,4537..5247)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      5745..5754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5935..5946
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    6159..6747
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6212..6232)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6630..6641)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7265..7445
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7516..8104)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7596..7615)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8275..9066,9067..9135))
                     /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     8898..8917
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9136..9240)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9315..9334)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgc
       61 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
      121 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
      181 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
      241 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
      301 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
      361 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcgcgttg acattgatta
      421 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      481 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      541 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      601 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      661 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      721 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      781 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      841 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      901 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      961 cgtgtacggt gggaggtcta tataagcagc gcgttttgcc tgtactgggt ctctctggtt
     1021 agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca
     1081 ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa
     1141 ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg gcgcccgaac
     1201 agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac tcggcttgct
     1261 gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa aattttgact
     1321 agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg ggggagaatt
     1381 agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata taaattaaaa
     1441 catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg cctgttagaa
     1501 acatcagaag gctgtagaca aatactggga cagctacaac catcccttca gacaggatca
     1561 gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca tcaaaggata
     1621 gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa caaaagtaag
     1681 accaccgcac agcaagcggc cggccgctga tcttcagacc tggaggagga gatatgaggg
     1741 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1801 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1861 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc
     1921 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1981 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     2041 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     2101 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     2161 aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga gaattaacaa
     2221 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     2281 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     2341 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     2401 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     2461 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     2521 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc
     2581 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg
     2641 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat
     2701 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc
     2761 cagtttggtt aattaagtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag
     2821 aagttggggg gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac
     2881 tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat
     2941 ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag
     3001 tctagagccg ccatggcggc aagggcaagc gtgctgagcg ggggggagct ggacaaatgg
     3061 gagaaaatcc ggctgagacc tggaggcaag aaacagtata aactgaagca catcgtgtgg
     3121 gctagcagag aactggagag gttcgcagtc aaccccggcc tgctggaaac atccgaggga
     3181 tgccgacaga ttctggggca gctgcagcct tccctgcaga ctggatccga ggaactgcgc
     3241 agtctgtaca acacagtggc caccctgtat tgcgtgcatc agcggattga cgtgaaagat
     3301 accaaggagg ctctggacaa aatcgaggaa gagcagaata agtccaagaa aaaggcacag
     3361 caggcggccg cagatacagg gaacaatagc caggtgtccc agaactaccc aatcgtccag
     3421 aatctgcagg gacagatggt gcaccaggcc atttctccca gaaccctgaa cgcttgggtg
     3481 aaagtggtcg aagagaaggc cttcagtcct gaagtcatcc caatgttttc tgctctgagt
     3541 gagggcgcaa cacctcaaga tctaaacaca atgctgaata ctgtgggcgg gcatcaggcc
     3601 gctatgcaga tgctgaagga aactatcaat gaagaggcag cagagtggga caggctgcac
     3661 ccagtgcatg caggaccaat tgcacctggg cagatgcgag aacctagagg atcagatatc
     3721 gccggcacca caagcacact gcaggagcag attgggtgga tgactcacaa cccccctata
     3781 ccggtaggag aaatctacaa aaggtggatc attctgggcc tgaataagat cgtccgcatg
     3841 tattcaccca ctagcatcct ggacattcga caaggcccaa aagagccctt ccgcgactac
     3901 gtggacagat tctacaagac cctgcgcgcc gaacaagctt cacaggaggt gaaaaactgg
     3961 atgactgaga ccctgctggt ccagaacgct aatcctgatt gcaaaacaat tctgaaggcc
     4021 ctggggccag gagctactct ggaagagatg atgaccgcat gtcagggagt gggaggacct
     4081 ggacacaagg ccagagtgct ggctgaggca atgagccagg tcaccaatcc agccacaatc
     4141 atgattcaga agggcaactt ccggaaccag cgcaaaaccg tgaagtgctt taactgtggc
     4201 aaagaggggc atatcgcaaa gaattgcagg gcccctcgga aaaagggatg ctggaaatgt
     4261 ggaaaggaag gccaccagat gaaggactgt actgagagac aggctaactt cctgggcaaa
     4321 atttggccct cccataaggg gagacccggg aattttctgc agagccggcc tgagccaacc
     4381 gcaccacccg aagagagttt caggtttggc gaagagacta ccacaccatc acagaaacag
     4441 gaacccatcg acaaggagct gtatcctctg gcaagtctgc gaagcctgtt tggaagcgac
     4501 ccctcatctc agggcggaag cggcggatcc atggtctcaa agggggaaga gctgtttact
     4561 ggggtcgtgc ctattctggt cgaactggac ggggatgtca acggccacaa gtttagtgtg
     4621 agcggagagg gagaaggcga cgccacttac ggaaagctga ccctgaaatt catttgcacc
     4681 acaggcaagc tgccagtgcc ctggcctacc ctggtcacta ccctgacata cggcgtccag
     4741 tgttttagta ggtatcctga ccacatgaag cagcatgatt tctttaaatc agctatgcca
     4801 gagggatacg tgcaggaacg caccattttc tttaaggacg atggcaacta caagactcga
     4861 gcagaggtga agttcgaagg ggacaccctg gtcaaccgga tcgagctgaa gggaatcgac
     4921 tttaaagaag atggaaacat cctgggccac aagctggagt acaactataa ttctcataac
     4981 gtgtacatca tggccgataa gcagaaaaac gggattaagg tcaatttcaa aatcagacac
     5041 aacatcgaag acggcagcgt gcagctggct gatcattacc agcagaacac accaattggc
     5101 gacgggccag tgctgctgcc agataatcac tatctgagca ctcagtccgc cctgtctaag
     5161 gaccctaacg agaaacgcga tcatatggtg ctgctggagt tcgtcacagc cgctgggatc
     5221 actctgggaa tggacgagct gtataaatga gaattctcga ccttgactgg cggcgcgacc
     5281 ttgaggcctg cgttcgcctc agttgccccc tctgtgcaat ggggagacgc gcctcatcgc
     5341 ttgacaacgg ccgaagagcc gccgcgcttc cgtctcccgc gtgcgcgcgc catgctgccc
     5401 acccccgttc cgcactgacc ctcccccgtg ccccgcgtcc cgtactgccg ccccgccccg
     5461 agtcccatgc cgcagccacc gcgacggagc ccgcaggcgg gaacctgcct ccgcgcgtta
     5521 gcgcgcacgc gcgcctcatg tgtcgtcccc atcagcgccg gcttccgtct ataggccaga
     5581 tgcactgtca ctctggcgaa gtcgcagacc cgattggccg ggacggaggc gcgagaccgg
     5641 gttgcgggcg gggccgaacg tggtataaaa ggggcgggag gccaggctcg tgccgttttg
     5701 cagacgccac cgccgaggaa aaccgtgtac tattagccac gcgtgccgcc atggctaaac
     5761 cactgtccca ggaagaatcc acactgatcg aacgagcaac cgctactatc aactcaatcc
     5821 caatctccga agactactct gtggcctccg ccgccctgag cagcgacgga cgaatcttca
     5881 ccggcgtgaa cgtctaccac tttaccggcg gcccctgcgc tgagctggtg gtcctgggaa
     5941 ctgcagccgc tgcagccgct ggaaatctga cctgtatcgt ggctatcggg aacgaaaata
     6001 ggggaatcct gtctccatgc ggccggtgta gacaggtcct gctggatctg catcccggga
     6061 tcaaggcaat tgtgaaagac tcagatggac agcccacagc cgtcggcatt cgcgagctgc
     6121 tgcctagcgg ctatgtgtgg gaagggtgag cgatcgctaa tcaacctctg gattacaaaa
     6181 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     6241 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     6301 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     6361 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     6421 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     6481 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     6541 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     6601 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     6661 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     6721 ggatctccct ttgggccgcc tccccgctta atcgcgtcga gacctagaaa aacatggagc
     6781 aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag aagcacaaga
     6841 ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa tgacttacaa
     6901 ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca
     6961 ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag gctacttccc
     7021 tgattggcag aactacacac cagggccagg gatcagatat ccactgacct ttggatggtg
     7081 ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag gagagaacac
     7141 ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag aagtattaga
     7201 gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc atccggactg
     7261 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
     7321 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
     7381 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
     7441 tagcagggcc cgtttcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7501 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7561 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7621 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7681 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7741 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7801 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7861 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7921 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     7981 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     8041 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     8101 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     8161 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     8221 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     8281 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8341 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8401 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8461 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8521 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8581 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8641 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8701 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8761 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8821 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8881 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8941 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     9001 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     9061 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     9121 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     9181 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     9241 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     9301 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     9361 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     9421 ggcttgaccg acaattgcat gaag
//
