LOCUS       sequence-14077-e        8604 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 8604)
  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 8604)
  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.hPGK.GP91
FEATURES             Location/Qualifiers
     source          1..8604
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..515
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     434..452
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      546..555
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2492..3080
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2545..2565)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2963..2974)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             3167..3400
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3472..3593
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3509..3528)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3563..3582
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3612..3747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3779..3798)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3780..3798)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3808..3825)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3808..3824)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3817..3839)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3969..4424
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4056..4075)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4266..4287
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4453..4557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4558..4626,4627..5418)
                     /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(4776..4795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5589..6177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6078..6097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6331..6348
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6465..6486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6501..6506,6507..6524,6525..6531)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6539..6555
                     /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     6544..6566
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6563..6579
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6563..6579
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6598..6618
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6600..6618
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             6874..7054
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7098..7223
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7716..7949
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             8134..8178
                     /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    8445..8562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ccacggggtt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct
       61 gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca
      121 cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc
      181 ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac
      241 aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg
      301 cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcgggg cgcgccgaga
      361 gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt
      421 tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct
      481 ccctcgttga ccgaatcacc gacctctctc cccaggggga tcccccgggg gatctatgag
      541 atcctgccac catggggaac tgggctgtga atgaggggct ctccattttt gtcattctgg
      601 tttggctggg gttgaacgtc ttcctctttg tctggtatta ccgggtttat gatattccac
      661 ctaagttctt ttacacaaga aaacttcttg ggtcagcact ggcactggcc agggcccctg
      721 cagcctgcct gaatttcaac tgcatgctga ttctcttgcc agtctgtcga aatctgctgt
      781 ccttcctcag gggttccagt gcgtgctgct caacaagagt tcgaagacaa ctggacagga
      841 atctcacctt tcataaaatg gtggcatgga tgattgcact tcactctgcg attcacacca
      901 ttgcacatct atttaatgtg gaatggtgtg tgaatgcccg agtcaataat tctgatcctt
      961 attcagtagc actctctgaa cttggagaca ggcaaaatga aagttatctc aattttgctc
     1021 gaaagagaat aaagaaccct gaaggaggcc tgtacctggc tgtgaccctg ttggcaggca
     1081 tcactggagt tgtcatcacg ctgtgcctca tattaattat cacttcctcc accaaaacca
     1141 tccggaggtc ttactttgaa gtcttttggt acacacatca tctctttgtg atcttcttca
     1201 ttggccttgc catccatgga gctgaacgaa ttgtacgtgg gcagaccgca gagagtttgg
     1261 ctgtgcataa tataacagtt tgtgaacaaa aaatctcaga atggggaaaa ataaaggaat
     1321 gcccaatccc tcagtttgct ggaaaccctc ctatgacttg gaaatggata gtgggtccca
     1381 tgtttctgta tctctgtgag aggttggtgc ggttttggcg atctcaacag aaggtggtca
     1441 tcaccaaggt ggtcactcac cctttcaaaa ccatcgagct acagatgaag aagaaggggt
     1501 tcaaaatgga agtgggacaa tacatttttg tcaagtgccc aaaggtgtcc aagctggagt
     1561 ggcacccttt tacactgaca tccgcccctg aggaagactt ctttagtatc catatccgca
     1621 tcgttgggga ctggacagag gggctgttca atgcttgtgg ctgtgataag caggagtttc
     1681 aagatgcgtg gaaactacct aagatagcgg ttgatgggcc ctttggcact gccagtgaag
     1741 atgtgttcag ctatgaggtg gtgatgttag tgggagcagg gattggggtc acacccttcg
     1801 catccattct caagtcagtc tggtacaaat attgcaataa cgccaccaat ctgaagctca
     1861 aaaagatcta cttctactgg ctgtgccggg acacacatgc ctttgagtgg tttgcagatc
     1921 tgctgcaact gctggagagc cagatgcagg aaaggaacaa tgccggcttc ctcagctaca
     1981 acatctacct cactggctgg gatgagtctc aggccaatca ctttgctgtg caccatgatg
     2041 aggagaaaga tgtgatcaca ggcctgaaac aaaagacttt gtatggacgg cccaactggg
     2101 ataatgaatt caagacaatt gcaagtcaac accctaatac cagaatagga gttttcctct
     2161 gtggacctga agccttggct gaaaccctga gtaaacaaag catctccaac tctgagtctg
     2221 gccctcgggg agtgcatttc attttcaaca aggaaaactt ctaacttgtc tcttccatga
     2281 ggaaataaat gtgggttgtg ctgccaaatg ctcaaataat gctaattgat aatataaata
     2341 ccccctgctt aaaaatggac aaaaagaaac tataatgtaa tggttttccc ttaaaggaat
     2401 gtcaaagatt gtttgatagt gataagttac atttatgtgg agctcgcccg ggctgcagga
     2461 attcgatatc aagcttatcg ataccgtcga caatcaacct ctggattaca aaatttgtga
     2521 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     2581 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     2641 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     2701 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     2761 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     2821 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     2881 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2941 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3001 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3061 cctttgggcc gcctccccgc ctggaattcg agctcggtac ctttaagacc aatgacttac
     3121 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
     3181 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
     3241 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
     3301 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
     3361 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
     3421 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
     3481 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3541 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     3601 tggctctagc tatcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3661 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3721 agtagtgagg aggctttttt ggaggcctag gcttttgcgt cgagacgtac ccaattcgcc
     3781 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     3841 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3901 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3961 atggcgcgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4021 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4081 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4141 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4201 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4261 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4321 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4381 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttcccagg tggcactttt
     4441 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4501 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4561 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4621 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4681 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4741 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4801 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4861 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4921 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4981 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5041 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5101 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5161 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5221 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5281 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5341 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5401 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5461 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5701 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5941 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6241 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6301 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6361 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6421 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6481 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6541 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
     6601 attaaccctc actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa
     6661 tactcttgta gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga
     6721 aaaagcaccg tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa
     6781 ggcaacagac gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata
     6841 ttgtatttaa gtgcctagct cgatacaata aacgggtctc tctggttaga ccagatctga
     6901 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
     6961 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
     7021 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacctgaaag
     7081 cgaaagggaa accagagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa
     7141 gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg
     7201 agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa
     7261 aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa
     7321 gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta
     7381 gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat
     7441 tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca
     7501 aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag
     7561 cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta
     7621 tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga
     7681 agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg
     7741 ggagcagcag gaagcactat gggcgcagcc tcaatgacgc tgacggtaca ggccagacaa
     7801 ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag
     7861 catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg
     7921 gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt
     7981 tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg
     8041 aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac
     8101 tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta
     8161 gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa
     8221 ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct
     8281 atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc
     8341 ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac
     8401 agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa aagaaaaggg
     8461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     8521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttatcgatca cgagactagc
     8581 ctcgagaagc ttgatatcga attc
//
