LOCUS       sequence-194236-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LTR drives GFP and internal SFFV promoter drives codon optimized tat
            and rev..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  AUTHORS   McCauley SM, Kim K, Nowosielska A, Dauphin A, Yurkovetskiy L, Diehl 
            WE, Luban J
  TITLE     Intron-containing RNA from the HIV-1 provirus activates type I 
            interferon and inflammatory cytokines.
  JOURNAL   Nat Commun. 2018 Dec 13;9(1):5305. doi: 10.1038/s41467-018-07753-2.
  PUBMED    30546110
REFERENCE   2  (bases 1 to 7126)
  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: "Nat 
            Commun."; date: "2018-12-13"; volume: "9(1):5305. doi"; pages: " 
            10.1038/s41467-018-07753-2"
COMMENT     Sequence Label: pALPS_3'LTR GFP@gag start SFFV tat-P2A-rev
FEATURES             Location/Qualifiers
     source          1..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(240..260)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        245..574
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      425..560
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     487..506
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        586..965
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        967..1165
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1120..1140
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1183..1363
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     CDS             join(1519..1521,1522..1524,1525..2235)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2398..2631
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2816..2860
                     /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    3158..3275
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3412..3819
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             4124..4180
                     /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"
     CDS             4403..4429
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     LTR             4673..5306
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5374..5962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5454..5473)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6133..6924,6925..6993))
                     /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     6756..6775
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6994..7098)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgctgtg
      241 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      301 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      361 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      421 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
      481 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
      541 aggaggcttt tttggaggcc taggcttttg caaaaagctc gcgttgacat tgattattga
      601 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      661 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      721 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      781 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      841 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      901 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      961 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     1021 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1081 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1141 tacggtggga ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac
     1201 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     1261 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     1321 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg
     1381 acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag
     1441 cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg
     1501 gaggctagaa ggagagagat ggtgtcaaaa ggagaagaac tgttcaccgg agtggtccca
     1561 atcctggtgg agctggacgg agatgtcaat ggccacaagt tcagcgtgag cggagaggga
     1621 gaaggagacg ctacatacgg caagctgact ctgaaattca tttgcaccac agggaagctg
     1681 cccgtgcctt ggccaaccct ggtcactacc ctgacatacg gcgtccagtg ttttagcaga
     1741 tatccagacc acatgaagca gcatgatttc tttaaatccg ccatgcccga gggctacgtg
     1801 caggaacgga caatcttctt taaggacgat ggaaactata aaactagagc tgaggtgaag
     1861 ttcgaaggag acaccctggt caacaggatc gagctgaagg gcattgactt taaagaagat
     1921 gggaatattc tgggacacaa gctggagtac aactacaact ctcataacgt gtatatcatg
     1981 gcagataagc agaaaaacgg cattaaggtc aatttcaaaa tccgccacaa tattgaagac
     2041 gggagcgtgc agctggccga tcattaccag cagaacacac ctatcggcga cggacccgtg
     2101 ctgctgcctg ataatcacta tctgtctact cagagtgccc tgtcaaagga cccaaacgag
     2161 aaacgagatc atatggtgct gctggagttt gtcaccgctg ctggaatcac actgggcatg
     2221 gatgagctgt acaagtaacg gccggccgct gatcttcaga cctggaggag gagatatgag
     2281 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     2341 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     2401 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     2461 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     2521 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     2581 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     2641 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     2701 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     2761 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     2821 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     2881 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     2941 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     3001 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     3061 aggtggagag agagacagag acagatccat tcgattagtg aacggatcgg cactgcgtgc
     3121 gccaattctg cagacaaatg gcagtattca tccacaattt taaaagaaaa ggggggattg
     3181 gggggtacag tgcaggggaa agaatagtag acataatagc aacagacata caaactaaag
     3241 aattacaaaa acaaattaca aaaattcaaa attttcgggt ttattacagg gacagcagag
     3301 atccagtttg gttaattaac tgcagccccg ataaaataaa agattttatt tagtctccag
     3361 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctgc agtaacgcca
     3421 ttttgcaagg catggaaaaa taccaaacca agaatagaga agttcagatc aagggcgggt
     3481 acatgaaaat agctaacgtt gggccaaaca ggatatctgc ggtgagcagt ttcggccccg
     3541 gcccggggcc aagaacagat ggtcaccgca gtttcggccc cggcccgagg ccaagaacag
     3601 atggtcccca gatatggccc aaccctcagc agtttcttaa gacccatcag atgtttccag
     3661 gctcccccaa ggacctgaaa tgaccctgcg ccttatttga attaaccaat cagcctgctt
     3721 ctcgcttctg ttcgcgcgct tctgcttccc gagctctata aaagagctca caacccctca
     3781 ctcggcgcgc cagtcctccg acagactgag tcgcccgggg gtctagaagc gctggatccg
     3841 tttaaacgcg gccgcgatgg aacccgtgga ccccagactg gaaccttgga aacaccccgg
     3901 atcacagcct aaaacagcct gcacaaactg ctactgcaaa aagtgctgtt tccactgcca
     3961 ggtgtgcttt atgactaagg ccctggggat ctcttacgga aggaagaaac ggagacagag
     4021 gcgccgagcc caccagaaca gtcagaccca tcaggctagc ctgtccaaac agccaacttc
     4081 tcagagtcgc ggcgacccca ccggacctaa ggagggatcc ggagcaacaa acttctcact
     4141 gctgaaacag gccggagatg tggaggaaaa tccagggccc atggccggaa ggtccggcga
     4201 ctctgatgag gaactgatcc gaacagtccg cctgattaag ctgctgtacc agtctaaccc
     4261 ccctccaaat cccgagggga ctcggcaggc tcgaagaaac aggcgccgac ggtggagaga
     4321 gaggcagcga cagatccatt caattagcga acggattctg agcacctatc tgggcagatc
     4381 cgcagaacct gtgccactgc agctgccccc tctggagaga ctgacactgg actgcaatga
     4441 agattgtggg accagcggaa cacagggcgt cggcagccct cagatcctgg tggagagccc
     4501 taccgtcctg gaaagcggaa caaaagagtg acccggggaa ttcacgcgta tcgataccgg
     4561 tggcccctgg ggccgcgatc gcagacctgc aggtggcctc aggtaccttt aagaccaatg
     4621 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     4681 ctaattcact cccaaagaag acaagatatc cttgatctgt ggatctacca cacacaaggc
     4741 tacttccctg attggcagaa ctacacacca gggccagggg tcagatatcc actgaccttt
     4801 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga
     4861 gagaacacca gcttgttaca ccctgtgagc ctgcatggaa tggatgaccc tgagagagaa
     4921 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat
     4981 ccggagtact tcaagaactg ctgacatcga gcttgctaca agggactttc cgctggggac
     5041 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atgctgcata
     5101 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
     5161 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     5221 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     5281 ttagtcagtg tggaaaatct ctagcaggcg tttcatgtga gcaaaaggcc agcaaaaggc
     5341 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5401 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5461 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5521 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5581 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5641 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5701 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5761 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5821 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5881 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5941 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6001 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6061 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6121 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6181 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6241 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6301 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6361 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6421 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6481 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6541 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6601 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6661 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6721 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6781 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6841 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6901 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6961 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7021 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7081 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac
//
