LOCUS       sequence-14072-e        7343 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7343)
  AUTHORS   Barde I, Laurenti E, Verp S, Wiznerowicz M, Offner S, Viornery A, 
            Galy A, Trumpp A, Trono D
  TITLE     Lineage- and stage-restricted lentiviral vectors for the gene 
            therapy of chronic granulomatous disease.
  JOURNAL   Gene Ther. 2011 Nov;18(11):1087-97. doi: 10.1038/gt.2011.65. Epub 
            2011 May 5.
  PUBMED    21544095
REFERENCE   2  (bases 1 to 7343)
  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/gt.2011.65"; journalName: "Gene Ther"; date: "2011-11"; 
            volume: "18"; issue: "11"; pages: "1087-97"
COMMENT     Sequence Label: pRRL.SP107.gp47.eGFP
FEATURES             Location/Qualifiers
     source          1..7343
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             408..452
                     /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    719..836
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      1336..1345
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1342..1344,1345..1347,1348..2058)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1342..1344,1345..1347,1348..2058)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1387..1408)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1648..1667)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1995..2016
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2110..2698
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2163..2183)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2581..2592)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2785..3018
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3090..3211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3127..3146)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3181..3200
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3230..3365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3397..3416)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3398..3416)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3426..3443)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3426..3442)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3435..3457)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3587..4042
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3674..3693)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3884..3905
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4071..4175
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4176..4244,4245..5036)
                     /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(4394..4413)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5207..5794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5696..5715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6080..6101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6116..6121,6122..6139,6140..6146)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6154..6170
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6159..6181
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6178..6194
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6178..6194
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6213..6233
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6215..6233
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6261..6487
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6488..6668
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6715..6840
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(7333..7343,1..223)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
       61 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
      121 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
      181 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
      241 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
      301 tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca
      361 caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag
      421 aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc
      481 tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt
      541 ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga
      601 cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag
      661 agagagacag agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt
      721 ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag
      781 caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa aattttatcg
      841 atcacgagac tagcctcgag aagcttgatg atccgtttaa ttcggtaccg agctcttacg
      901 cgtgctagcg agggcggaca agcgacttcc tctttccagc agaaaaggag aagtaggagc
      961 caagatttcc aaactctgtg gttgccttga agcgacttcc tctttccaga agcgacttcc
     1021 tctttccagc agaaaaggag aagtaggagc tcctacttct ccttttctgc caagatttcc
     1081 aaactctgtg gttgccttgc tcctacttct ccttttctgc agaaaaggag aagtaggagc
     1141 tggaaagagg aagtcgcttg agggcggacg ctagcccggg ctcgagtacc ctccctgccc
     1201 cctgtcccga ccgcgacaaa agcgacttcc tctttccagt gcatttaagg cgcagcctgg
     1261 aagtgccagg gagcactgga ggccacccag taccgggatc caggcctaag cttacgcgtc
     1321 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1381 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1441 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1501 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1561 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1621 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1681 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1741 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1801 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1861 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1921 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1981 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2041 atggacgagc tgtacaagtc cggactcaga tctcgactag ctagtagcta gctagaaacg
     2101 ctggtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2161 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2221 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     2281 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     2341 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     2401 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     2461 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     2521 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2581 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2641 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     2701 ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     2761 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc
     2821 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     2881 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     2941 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     3001 tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat ttataacttg
     3061 caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat aatggttaca
     3121 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3181 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta tcccgcccct
     3241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3301 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3361 aggcctaggc ttttgcgtcg agacgtaccc aattcgccct atagtgagtc gtattacgcg
     3421 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     3481 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     3541 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc gccctgtagc
     3601 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3661 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3721 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3781 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3841 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3901 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3961 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     4021 aaaatattaa cgtttacaat ttcccaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4081 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4141 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4201 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4261 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4321 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4381 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4441 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4501 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4561 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4621 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4681 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4741 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4801 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4861 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4921 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4981 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     5041 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5101 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5161 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5221 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5281 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5341 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5401 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5461 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5521 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5581 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5641 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5701 acgcctggta tctttatagt cctgtcgggt ttcgcgcacc tctgacttga gcgtcgattt
     5761 ttgtgatgct cgtcaggggg gcgcgtgatg gaaaaacgcc agcaacgcgc ctttttacgg
     5821 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5881 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5941 gagcgcacag gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     6001 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6061 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     6121 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     6181 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     6241 agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt gcaacatggt
     6301 aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat gccgattggt
     6361 ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc tgacatggat
     6421 tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc tagctcgata
     6481 cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6541 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6601 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6661 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     6721 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     6781 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     6841 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     6901 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     6961 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     7021 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     7081 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     7141 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
     7201 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
     7261 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
     7321 agcagtggga ataggagctt tgt
//
