LOCUS       sequence-45368-e        9310 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 9310)
  AUTHORS   Astrakhan A, Sather BD, Ryu BY, Khim S, Singh S, Humblet-Baron S, 
            Ochs HD, Miao CH, Rawlings DJ
  TITLE     Ubiquitous high-level gene expression in hematopoietic lineages 
            provides effective lentiviral gene therapy of murine Wiskott-Aldrich
            Syndrome.
  JOURNAL   Blood. 2012 Mar 19.
  PUBMED    22431569
REFERENCE   2  (bases 1 to 9310)
  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: "Blood. 2012
            Mar 19."
COMMENT     Sequence Label: pRRL WS1.6 WASp
FEATURES             Location/Qualifiers
     source          1..9310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             236..416
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1924
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_feature    5164..5752
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5217..5237)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5635..5646)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5839..6072
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6144..6265
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6181..6200)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6235..6254
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6284..6419
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6451..6470)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6452..6470)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6480..6497)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6480..6496)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6489..6511)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6641..7096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6728..6747)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6938..6959
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        7125..7229
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7230..7298,7299..8090)
                     /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(7448..7467)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8261..8849
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8750..8769
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9003..9020
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9137..9158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9173..9178,9179..9196,9197..9203)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9211..9227
                     /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     9216..9238
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9235..9251
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9235..9251
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     9270..9290
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        9272..9290
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag
       61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg
      121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact
      181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacaa taaacgggtc
      241 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
      301 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
      361 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtgg
      421 cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccgct gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cctcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa
     1921 ttttatcgat cacgagacta gcctcgagaa gcttgatatc gaattccacg gggttggacg
     1981 cgttgcagtg ttgctcccaa cagcaagaac cagaagcagc ccaaagggct gttacaggag
     2041 aatatggaca cccaggctgc acatgcacac catggaatgc tgtatggcag tggaaataaa
     2101 tgaacagcta ccactatagg caaacaggaa tcacagcaac agccaagagt gaaggcgtgg
     2161 agggacgaga ccatgcactc acacctggcc tgcctggctc gcactccggg caaaggggtc
     2221 agaacagtga ctggcacaca cgttaagtgc tatgtgagtg ttaagataaa actaggatgt
     2281 ccagtgggga gaaagcaagc ctttgaagat tatgtgcttt tacaaacttc aagtgcaatg
     2341 aaaactaaac aagatgttgt tcaggcattc atatatgata taaagttcct ttctttaaaa
     2401 aagggatggg ctgggcacgg tggctcacgc ctgtaattct aatactttgg gaggccgagg
     2461 caggtggatc acgaggtcga gaaatcgaga ccatcctggc caacatggtg aaaccctgtc
     2521 tctactaaaa atacaaaaaa attagctggg cgtggtggcg tgtgcctgta gtcccagcta
     2581 cttgggaggc tgaggcagga gagtcacttg aacccgggag gcaaaggttg cagtgagccg
     2641 agatcgtgcc acccgcactc cagcctggcg acagagtgag actccatctc aaaaaaaaaa
     2701 agaaaaaaaa aagtatgaca agcagaaagt aatttgggag ctgcggggag gcaagggtaa
     2761 gggatgggga agtggaccag aggcatatgc gtcattggca gtgtctaagc actcacgata
     2821 ggcgtggatc acaggggctc gctctgtaat taaaaggaaa agggtttttg ttgtgttgtt
     2881 gttgttgctg tttttgagac aagggtcttg ctctgtcatc atccaggctg gagtgcagtg
     2941 gtgcagtctc agctcactgc aacctccgcc tcctgggttc aagcgattct cctgcctcag
     3001 cctcctgagc agctaggact acaggtgtgt gccaccatgc ctggctaatt tttgtatttt
     3061 ttagtggaaa tggggttttg ccatgttgcc caggctcgtc ttgaactcct gacctcaagt
     3121 gatccactcg tctcggcctc ccaaagtgct gggattacag gtgtgagcta ttgtccccag
     3181 ccaaaaggaa aagttttact gtagtaaccc ttccggacta gggacctcgg gcctcagcct
     3241 caggctacct aggtgcttta gaaaggaggc cacccaggcc catgactact ccttgccaca
     3301 gggagccccg cacacagatg tgctaagctc tcgctgccag ccagagggag gagggtctga
     3361 gccagtcaga aggagatggg ccccagagag taagaaaggg ggaggaggac ccaagctgat
     3421 ccaaaaggtg ggtctaagca gtcaagtgga ggagggttcc aatctgatgg cggagggccc
     3481 aagctcagcc taacgaggag gccaggccca ccaaggggcc cctggaggac ttgtttccct
     3541 tgtcccttgt ggttttttgc atttcctgtt cccttgctgc tcattgcgga agttcctctt
     3601 cttaccctgc acccagagcc tcgccaggat ccagaagcca ccatgagtgg gggcccaatg
     3661 ggaggaaggc ccgggggccg aggagcacca gcggttcagc agaacatacc ctccaccctc
     3721 ctccaggacc acgagaacca gcgactcttt gagatgcttg gacgaaaatg cttgacgctg
     3781 gccactgcag ttgttcagct gtacctggcg ctgccccctg gagctgagca ctggaccaag
     3841 gagcattgtg gggctgtgtg cttcgtgaag gataaccccc agaagtccta cttcatccgc
     3901 ctttacggcc ttcaggctgg tcggctgctc tgggaacagg agctgtactc acagcttgtc
     3961 tactccaccc ccaccccctt cttccacacc ttcgctggag atgactgcca agcggggctg
     4021 aactttgcag acgaggacga ggcccaggcc ttccgggccc tcgtgcagga gaagatacaa
     4081 aaaaggaatc agaggcaaag tggagacaga cgccagctac ccccaccacc aacaccagcc
     4141 aatgaagaga gaagaggagg gctcccaccc ctgcccctgc atccaggtgg agaccaagga
     4201 ggccctccag tgggtccgct ctccctgggg ctggcgacag tggacatcca gaaccctgac
     4261 atcacgagtt cacgataccg tgggctccca gcacctggac ctagcccagc tgataagaaa
     4321 cgctcaggga agaagaagat cagcaaagct gatattggtg cacccagtgg attcaagcat
     4381 gtcagccacg tggggtggga cccccagaat ggatttgacg tgaacaacct cgacccagat
     4441 ctgcggagtc tgttctccag ggcaggaatc agcgaggccc agctcaccga cgccgagacc
     4501 tctaaactta tctacgactt cattgaggac cagggtgggc tggaggctgt gcggcaggag
     4561 atgaggcgcc aggagccact tccgccgccc ccaccgccat ctcgaggagg gaaccagctc
     4621 ccccggcccc ctattgtggg gggtaacaag ggtcgttctg gtccactgcc ccctgtacct
     4681 ttggggattg ccccaccccc accaacaccc cggggacccc cacccccagg ccgagggggc
     4741 cctccaccac caccccctcc agctactgga cgttctggac cactgccccc tccaccccct
     4801 ggagctggtg ggccacccat gccaccacca ccgccaccac cgccaccgcc gcccagctcc
     4861 gggaatggac cagcccctcc cccactccct cctgctctgg tgcctgccgg gggcctggcc
     4921 cctggtgggg gtcggggagc gcttttggat caaatccggc agggaattca gctgaacaag
     4981 acccctgggg ccccagagag ctcagcgctg cagccaccac ctcagagctc agagggactg
     5041 gtgggggccc tgatgcacgt gatgcagaag agaagcagag ccatccactc ctccgacgaa
     5101 ggggaggacc aggctggcga tgaagatgaa gatgatgaat gggatgactg aactagtgtc
     5161 gacaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5221 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5281 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5341 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5401 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5461 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5521 ctgttgggca ctgacaattc cgtggtgttg tcggggaagc tgacgtcctt tccatggctg
     5581 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     5641 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     5701 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggaatt
     5761 cgagctcggt acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt
     5821 taaaagaaaa ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt
     5881 ttgcttgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     5941 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     6001 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     6061 aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga
     6121 aatgaatatc agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa
     6181 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     6241 tgtccaaact catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc
     6301 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     6361 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     6421 aggcttttgc gtcgagacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca
     6481 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     6541 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     6601 ccttcccaac agttgcgcag cctgaatggc gaatggcgcg acgcgccctg tagcggcgca
     6661 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6721 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6781 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6841 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6901 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6961 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     7021 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     7081 ttaacgttta caatttccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     7141 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     7201 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     7261 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     7321 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     7381 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     7441 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     7501 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     7561 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     7621 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     7681 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     7741 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     7801 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     7861 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     7921 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     7981 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     8041 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     8101 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     8161 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     8221 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     8281 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     8341 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     8401 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     8461 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     8521 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     8581 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     8641 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     8701 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     8761 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     8821 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     8881 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     8941 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     9001 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     9061 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     9121 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     9181 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     9241 acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg gaacaaaagc
     9301 tggagctgca
//
