LOCUS       sequence-45127-e        8134 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 8134)
  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 8134)
  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 MND WASp
FEATURES             Location/Qualifiers
     source          1..8134
                     /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"
     primer_bind     2320..2344
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     misc_feature    3988..4576
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4041..4061)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4459..4470)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4663..4896
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4968..5089
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5005..5024)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5059..5078
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5108..5243
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5275..5294)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5276..5294)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5304..5321)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5304..5320)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5313..5335)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5465..5920
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5552..5571)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5762..5783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5949..6053
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6054..6122,6123..6914)
                     /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(6272..6291)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7085..7673
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7574..7593
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7827..7844
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7961..7982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7997..8002,8003..8020,8021..8027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8035..8051
                     /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     8040..8062
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8059..8075
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8059..8075
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8094..8114
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8096..8114
                     /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 gaattcccac ggggttggac
     1981 gcgtcttaat taaggatccg aacagagaga cagcagaata tgggccaaac aggatatctg
     2041 tggtaagcag ttcctgcccc gctcagggcc aagaacagtt ggaacagcag aatatgggcc
     2101 aaacaggata tctgtggtaa gcagttcctg ccccgctcag ggccaagaac agatggtccc
     2161 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     2221 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     2281 tgttcgcgcg cttctgctcc ccgagctcta tataagcaga gctcgtttag tgaaccgtca
     2341 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gactctagag
     2401 gatcgatccc ccgggctgca ggaattcaag cgagaagaca agggcagaaa gcaccatgag
     2461 tgggggccca atgggaggaa ggcccggggg ccgaggagca ccagcggttc agcagaacat
     2521 accctccacc ctcctccagg accacgagaa ccagcgactc tttgagatgc ttggacgaaa
     2581 atgcttgacg ctggccactg cagttgttca gctgtacctg gcgctgcccc ctggagctga
     2641 gcactggacc aaggagcatt gtggggctgt gtgcttcgtg aaggataacc cccagaagtc
     2701 ctacttcatc cgcctttacg gccttcaggc tggtcggctg ctctgggaac aggagctgta
     2761 ctcacagctt gtctactcca cccccacccc cttcttccac accttcgctg gagatgactg
     2821 ccaagcgggg ctgaactttg cagacgagga cgaggcccag gccttccggg ccctcgtgca
     2881 ggagaagata caaaaaagga atcagaggca aagtggagac agacgccagc tacccccacc
     2941 accaacacca gccaatgaag agagaagagg agggctccca cccctgcccc tgcatccagg
     3001 tggagaccaa ggaggccctc cagtgggtcc gctctccctg gggctggcga cagtggacat
     3061 ccagaaccct gacatcacga gttcacgata ccgtgggctc ccagcacctg gacctagccc
     3121 agctgataag aaacgctcag ggaagaagaa gatcagcaaa gctgatattg gtgcacccag
     3181 tggattcaag catgtcagcc acgtggggtg ggacccccag aatggatttg acgtgaacaa
     3241 cctcgaccca gatctgcgga gtctgttctc cagggcagga atcagcgagg cccagctcac
     3301 cgacgccgag acctctaaac ttatctacga cttcattgag gaccagggtg ggctggaggc
     3361 tgtgcggcag gagatgaggc gccaggagcc acttccgccg cccccaccgc catctcgagg
     3421 agggaaccag ctcccccggc cccctattgt ggggggtaac aagggtcgtt ctggtccact
     3481 gccccctgta cctttgggga ttgccccacc cccaccaaca ccccggggac ccccaccccc
     3541 aggccgaggg ggccctccac caccaccccc tccagctact ggacgttctg gaccactgcc
     3601 ccctccaccc cctggagctg gtgggccacc catgccacca ccaccgccac caccgccacc
     3661 gccgcccagc tccgggaatg gaccagcccc tcccccactc cctcctgctc tggtgcctgc
     3721 cgggggcctg gcccctggtg ggggtcgggg agcgcttttg gatcaaatcc ggcagggaat
     3781 tcagctgaac aagacccctg gggccccaga gagctcagcg ctgcagccac cacctcagag
     3841 ctcagaggga ctggtggggg ccctgatgca cgtgatgcag aagagaagca gagccatcca
     3901 ctcctccgac gaaggggagg accaggctgg cgatgaagat gaagatgatg aatgggatga
     3961 ctgagtggct gagttactag tgtcgacaat caacctctgg attacaaaat ttgtgaaaga
     4021 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     4081 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     4141 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     4201 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     4261 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     4321 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     4381 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     4441 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     4501 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     4561 tgggccgcct ccccgcctgg aattcgagct cggtaccttt aagaccaatg acttacaagg
     4621 cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact
     4681 cccaacgaag acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct
     4741 gagcctggga gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc
     4801 cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc
     4861 tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta
     4921 ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac ttgtttattg
     4981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctggc
     5101 tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     5161 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     5221 gtgaggaggc ttttttggag gcctaggctt ttgcgtcgag acgtacccaa ttcgccctat
     5281 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     5341 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     5401 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     5461 cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5521 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5581 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5641 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5701 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5761 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5821 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     5881 tttaacgcga attttaacaa aatattaacg tttacaattt cccaggtggc acttttcggg
     5941 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     6001 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     6061 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     6121 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     6181 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     6241 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     6301 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     6361 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     6421 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     6481 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     6541 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     6601 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6661 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6721 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6781 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6841 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6901 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6961 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     7021 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     7081 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     7141 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     7201 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     7261 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     7321 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     7381 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     7441 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     7501 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     7561 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     7621 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7681 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7741 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7801 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7861 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7921 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7981 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     8041 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     8101 accctcacta aagggaacaa aagctggagc tgca
//
