LOCUS       sequence-191197-        6683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  LTR drives GFP and internal SFFV promoter drives codon optimized 
            tat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6683)
  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 6683)
  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
FEATURES             Location/Qualifiers
     source          1..6683
                     /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        582..961
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        963..1161
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1116..1136
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             1179..1359
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     CDS             join(1515..1517,1518..1520,1521..2231)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    2394..2627
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2812..2856
                     /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    3154..3271
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3408..3815
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3859..3868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     LTR             4230..4863
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4931..5519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5011..5030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5690..6481,6482..6550))
                     /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     6313..6332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6551..6655)
                     /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 caaaaagctt tgacattgat tattgactag
      601 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      661 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      721 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      781 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      841 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      901 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      961 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1021 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1081 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1141 gtgggaggtc tatataagca gcgcgttttg cctgtactgg gtctctctgg ttagaccaga
     1201 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     1261 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     1321 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tggcgcccga acagggactt
     1381 gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg actcggcttg ctgaagcgcg
     1441 cacggcaaga ggcgaggggc ggcgactggt gagtacgcca aaaattttga ctagcggagg
     1501 ctagaaggag agagatggtg tcaaaaggag aagaactgtt caccggagtg gtcccaatcc
     1561 tggtggagct ggacggagat gtcaatggcc acaagttcag cgtgagcgga gagggagaag
     1621 gagacgctac atacggcaag ctgactctga aattcatttg caccacaggg aagctgcccg
     1681 tgccttggcc aaccctggtc actaccctga catacggcgt ccagtgtttt agcagatatc
     1741 cagaccacat gaagcagcat gatttcttta aatccgccat gcccgagggc tacgtgcagg
     1801 aacggacaat cttctttaag gacgatggaa actataaaac tagagctgag gtgaagttcg
     1861 aaggagacac cctggtcaac aggatcgagc tgaagggcat tgactttaaa gaagatggga
     1921 atattctggg acacaagctg gagtacaact acaactctca taacgtgtat atcatggcag
     1981 ataagcagaa aaacggcatt aaggtcaatt tcaaaatccg ccacaatatt gaagacggga
     2041 gcgtgcagct ggccgatcat taccagcaga acacacctat cggcgacgga cccgtgctgc
     2101 tgcctgataa tcactatctg tctactcaga gtgccctgtc aaaggaccca aacgagaaac
     2161 gagatcatat ggtgctgctg gagtttgtca ccgctgctgg aatcacactg ggcatggatg
     2221 agctgtacaa gtaacggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac
     2281 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
     2341 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
     2401 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
     2461 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
     2521 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
     2581 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
     2641 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
     2701 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
     2761 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
     2821 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
     2881 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
     2941 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
     3001 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
     3061 ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact gcgtgcgcca
     3121 attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg ggattggggg
     3181 gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa ctaaagaatt
     3241 acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca gcagagatcc
     3301 agtttggtta attaactgca gccccgataa aataaaagat tttatttagt ctccagaaaa
     3361 aggggggaat gaaagacccc acctgtaggt ttggcaagct agctgcagta acgccatttt
     3421 gcaaggcatg gaaaaatacc aaaccaagaa tagagaagtt cagatcaagg gcgggtacat
     3481 gaaaatagct aacgttgggc caaacaggat atctgcggtg agcagtttcg gccccggccc
     3541 ggggccaaga acagatggtc accgcagttt cggccccggc ccgaggccaa gaacagatgg
     3601 tccccagata tggcccaacc ctcagcagtt tcttaagacc catcagatgt ttccaggctc
     3661 ccccaaggac ctgaaatgac cctgcgcctt atttgaatta accaatcagc ctgcttctcg
     3721 cttctgttcg cgcgcttctg cttcccgagc tctataaaag agctcacaac ccctcactcg
     3781 gcgcgccagt cctccgacag actgagtcgc ccgggggtct agaagcgctg gatccgttta
     3841 aacgcggccg ccacgcgtgc caccatggag cccgtggacc ctcggctgga accttggaag
     3901 cacccagggt ctcagcccaa aaccgcatgc acaaactgct actgtaagaa atgctgtttc
     3961 cattgccagg tctgttttat gacaaaggcc ctgggcatct catatgggag aaagaaacgg
     4021 agacagaggc gccgagccca ccagaatagc cagacccatc aggctagcct gtccaagcag
     4081 cctacttctc agagtcgggg agatccaacc ggccccaaag agtgacccgg ggcgatcgca
     4141 gacctgcagg tggcctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
     4201 tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc aaagaagaca
     4261 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt ggcagaacta
     4321 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
     4381 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
     4441 tgtgagcctg catggaatgg atgaccctga gagagaagtg ttagagtgga ggtttgacag
     4501 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
     4561 acatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
     4621 cgggactggg gagtggcgag ccctcagatg ctgcatataa gcagctgctt tttgcctgta
     4681 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
     4741 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
     4801 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
     4861 gcaggcgttt catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4921 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4981 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     5041 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5101 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5161 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5221 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5281 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5341 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5401 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5461 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5521 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5581 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5641 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5701 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5761 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5821 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5881 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5941 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6001 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6061 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6121 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6181 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6241 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6301 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6361 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6421 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6481 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6541 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6601 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6661 ttccccgaaa agtgccacct gac
//
