LOCUS       sequence-14075-e        7353 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 7353)
  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 7353)
  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.SP146.gp47.eGFP
FEATURES             Location/Qualifiers
     source          1..7353
                     /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      1346..1355
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1352..1354,1355..1357,1358..2068)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1352..1354,1355..1357,1358..2068)
                     /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(1397..1418)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1658..1677)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2005..2026
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2120..2708
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2173..2193)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2591..2602)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2795..3028
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3100..3221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3137..3156)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3191..3210
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3240..3375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3407..3426)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3408..3426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3436..3453)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3436..3452)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3445..3467)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3597..4052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3684..3703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3894..3915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4081..4185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4186..4254,4255..5046)
                     /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(4404..4423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5217..5804
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5706..5725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    6090..6111
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6126..6131,6132..6149,6150..6156)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6164..6180
                     /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     6169..6191
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6188..6204
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6188..6204
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6223..6243
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6225..6243
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6271..6497
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6498..6678
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6725..6850
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(7343..7353,1..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 agggcggacc agaaaaggag aagtaggagc caagatttcc aaactctgtg
      961 gttgccttgc caagatttcc aaactctgtg gttgccttgc agaaaaggag aagtaggaga
     1021 agcgacttcc tctttccaga agcgacttcc tctttccaga ggaagagggc ggaggctcac
     1081 aaggcaacca cagagtttgg aaatcttgga agcgacttcc tctttccagc agaaaaggag
     1141 aagtaggaga agcgacttcc tctttccagg tccgccctcg ctagcccggg ctcgagtacc
     1201 ctccctgccc cctgtcccga ccgcgacaaa agcgacttcc tctttccagt gcatttaagg
     1261 cgcagcctgg aagtgccagg gagcactgga ggccacccag taccgggatc caggcctaag
     1321 cttacgcgtc ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac
     1381 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     1441 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     1501 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     1561 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     1621 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     1681 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     1741 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     1801 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     1861 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1921 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     1981 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2041 cactctcggc atggacgagc tgtacaagtc cggactcaga tctcgactag ctagtagcta
     2101 gctagaaacg ctggtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2161 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2221 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2281 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2341 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2401 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2461 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     2521 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2581 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2641 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2701 ctccccgcct ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta
     2761 gatcttagcc actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga
     2821 agacaagatc tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg
     2881 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     2941 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     3001 ttttagtcag tgtggaaaat ctctagcagt agtagttcat gtcatcttat tattcagtat
     3061 ttataacttg caaagaaatg aatatcagag agtgagagga acttgtttat tgcagcttat
     3121 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3181 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gctctagcta
     3241 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3301 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3361 gcttttttgg aggcctaggc ttttgcgtcg agacgtaccc aattcgccct atagtgagtc
     3421 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     3481 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     3541 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc
     3601 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3661 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3721 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3781 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3841 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3901 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3961 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     4021 gaattttaac aaaatattaa cgtttacaat ttcccaggtg gcacttttcg gggaaatgtg
     4081 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4141 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     4201 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4261 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4321 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4381 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4441 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4501 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4561 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4621 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4681 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4741 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4801 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4861 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4921 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4981 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5041 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5101 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5161 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5221 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5281 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5341 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     5401 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     5461 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     5521 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     5581 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     5641 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     5701 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgcgcacc tctgacttga
     5761 gcgtcgattt ttgtgatgct cgtcaggggg gcgcgtgatg gaaaaacgcc agcaacgcgc
     5821 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5881 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5941 ccgaacgacc gagcgcacag gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     6001 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     6061 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     6121 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     6181 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa
     6241 agggaacaaa agctggagct gcaagcttaa tgtagtctta tgcaatactc ttgtagtctt
     6301 gcaacatggt aacgatgagt tagcaacatg ccttacaagg agagaaaaag caccgtgcat
     6361 gccgattggt ggaagtaagg tggtacgatc gtgccttatt aggaaggcaa cagacgggtc
     6421 tgacatggat tggacgaacc actgaattgc cgcattgcag agatattgta tttaagtgcc
     6481 tagctcgata cataaacggg tctctctggt tagaccagat ctgagcctgg gagctctctg
     6541 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag
     6601 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag
     6661 tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga
     6721 ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg
     6781 gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg
     6841 cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag
     6901 gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga
     6961 acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg
     7021 acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt
     7081 agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga
     7141 caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct
     7201 tcagacctgg aggaggagat atgagggaca attggagaag tgaattatat aaatataaag
     7261 tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga
     7321 gagaaaaaag agcagtggga ataggagctt tgt
//
