LOCUS       sequence-194210-        7413 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes eGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7413)
  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 7413)
  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 eGFP
FEATURES             Location/Qualifiers
     source          1..7413
                     /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"
     misc_feature    1410..1535
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2032..2265
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2450..2494
                     /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    2792..2909
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3046..3453
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     regulatory      3487..3496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3493..3495,3496..3498,3499..4212)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3493..3495,3496..3498,3499..4209)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3538..3559)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3799..3818)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4146..4167
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4304..4892
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4357..4377)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4775..4786)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5410..5590
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(5661..6249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5741..5760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6420..7211,7212..7280))
                     /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     7043..7062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7281..7385)
                     /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 gggtgcgaga gcgtcagtat taagcggggg agaattagat
     1561 cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata
     1621 tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat
     1681 cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag
     1741 aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga
     1801 taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca
     1861 ccgcacagca agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa
     1921 ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc
     1981 caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt
     2041 gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac
     2101 ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt tgctgagggc
     2161 tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc agctccaggc
     2221 aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg
     2281 ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc
     2341 tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta
     2401 cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca
     2461 agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg
     2521 gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt
     2581 ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca
     2641 gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg
     2701 agagagagac agagacagat ccattcgatt agtgaacgga tcggcactgc gtgcgccaat
     2761 tctgcagaca aatggcagta ttcatccaca attttaaaag aaaagggggg attggggggt
     2821 acagtgcagg ggaaagaata gtagacataa tagcaacaga catacaaact aaagaattac
     2881 aaaaacaaat tacaaaaatt caaaattttc gggtttatta cagggacagc agagatccag
     2941 tttggttaat taactgcagc cccgataaaa taaaagattt tatttagtct ccagaaaaag
     3001 gggggaatga aagaccccac ctgtaggttt ggcaagctag ctgcagtaac gccattttgc
     3061 aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc gggtacatga
     3121 aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc cccggcccgg
     3181 ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga acagatggtc
     3241 cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt ccaggctccc
     3301 ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct gcttctcgct
     3361 tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc ctcactcggc
     3421 gcgccagtcc tccgacagac tgagtcgccc gggggtctag aagcgctgga tccccgggta
     3481 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     3541 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     3601 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     3661 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     3721 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     3781 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3841 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3901 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3961 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     4021 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     4081 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     4141 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     4201 ctgtacaagt aaagcggccg cccagcacag tggctcgagc cgcgggttaa ctggccagaa
     4261 ttcacgcgta tcgataccgg tggcccctgg ggccgcgatc gctaatcaac ctctggatta
     4321 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4381 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4441 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4501 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4561 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4621 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4681 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     4741 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4801 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4861 gagtcggatc tccctttggg ccgcctcccc gcttaatcgc gtcgagacct agaaaaacat
     4921 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     4981 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     5041 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     5101 attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac acaaggctac
     5161 ttccctgatt ggcagaacta cacaccaggg ccagggatca gatatccact gacctttgga
     5221 tggtgctaca agctagtacc agttgagcaa gagaaggtag aagaagccaa tgaaggagag
     5281 aacacccgct tgttacaccc tgtgagcctg catgggatgg atgacccgga gagagaagta
     5341 ttagagtgga ggtttgacag ccgcctagca tttcatcaca tggcccgaga gctgcatccg
     5401 gactgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta
     5461 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc
     5521 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa
     5581 atctctagca gggcccgttt catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5821 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6061 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6361 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6421 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6481 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6541 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6601 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6661 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6721 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6781 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6841 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6901 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6961 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7021 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7081 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7141 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7201 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7261 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7321 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7381 ccgcgcacat ttccccgaaa agtgccacct gac
//
