LOCUS       sequence-469967-        8373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible lentiviral expression of ITPA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8373)
  AUTHORS   Tang S, Stokasimov E, Cui Y, Pellman D
  TITLE     Breakage of cytoplasmic chromosomes by pathological DNA base 
            excision repair.
  JOURNAL   Nature. 2022 Jun;606(7916):930-936. doi: 10.1038/s41586-022-04767-1.
            Epub 2022 Apr 27.
  PUBMED    35477155
REFERENCE   2  (bases 1 to 8373)
  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/s41586-022-04767-1"; journalName: "Nature"; date: 
            "2022-06"; volume: "606"; issue: "7916"; pages: "930-936"
COMMENT     Sequence Label: pHAGE Flag HA ITPA
FEATURES             Location/Qualifiers
     source          1..8373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1000..1021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1036..1041,1042..1059,1060..1066)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1074..1090
                     /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     1079..1101
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1098..1114
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1098..1114
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1164..1183
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             1375..2008
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    2055..2180
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2677..2910
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             3095..3139
                     /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    3436..3553
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3605..3908
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3909..4112
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4063..4083
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4109..4133
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      4209..4218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4218..4241
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4254..4280
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    4296..4320
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4903..4927)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    4940..5507
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4957..5509
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5101..5118)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5328..5347
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5521..6120
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5521..5540)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     6017..6037
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     misc_feature    6137..6725
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6190..6210)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6608..6619)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6800..7033
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(7152..7170)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7238..7342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7343..7411,7412..8203)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7561..7580)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcaagc tcatggctga
      841 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      901 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctccccgt ggcacgacag
      961 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     1021 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     1081 cggataacaa tttcacacag gaaacagcta tgacatgatt acgaatttca caaataaagc
     1141 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     1201 ctggatcaac tggataactc aagctaacca aaatcatccc aaacttccca ccccataccc
     1261 tattaccact gccaattacc tgtggtttca tttactctaa acctgtgatt cctctgaatt
     1321 attttcattt taaagaaatt gtatttgtta aatatgtact acaaacttag tagttggaag
     1381 ggctaattca ctcccaaaga agacaagata tccttgatct gtggatctac cacacacaag
     1441 gctacttccc tgattagcag aactacacac cagggccagg ggtcagatat ccactgacct
     1501 ttggatggtg ctacaagcta gtaccagttg agccagataa ggtagaagag gccaataaag
     1561 gagagaacac cagcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     1621 aagtgttaga gtggaggttt gacagccgcc tagcatttca tcacgtggcc cgagagctgc
     1681 atccggagta cttcaagaac tgctgatatc gagcttgcta caagggactt tccgctgggg
     1741 actttccagg gaggcgtggc ctgggcggga ctggggagtg gcgagccctc agatcctgca
     1801 tataagcagc tgctttttgc ctgtactggg tctctctggt tagaccagat ctgagcctgg
     1861 gagctctctg gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg
     1921 cttcaagtag tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc
     1981 ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag
     2041 ggaaaccaga ggagctctct cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag
     2101 gcgaggggcg gcgactggtg agtacgccaa aaattttgac tagcggaggc tagaaggaga
     2161 gagatgggtg cgagagcgtc agtattaagc gggggagaat tagatcgcga tgggaaaaaa
     2221 ttcggttaag gccaggggga aagaaaaaat ataaattaaa acatatagta tgggcaagca
     2281 gggagctaga acgattcgca gttaatcctg gcctgttaga aacatcagaa ggctgtagac
     2341 aaatactggg acagctacaa ccatcccttc agacaggatc agaagaactt agatcattat
     2401 ataatacagt agcaaccctc tattgtgtgc atcaaaggat agagataaaa gacaccaagg
     2461 aagctttaga caagatagag gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg
     2521 ccggccgctg atcttcagac ctggaggagg agatatgagg gacaattgga gaagtgaatt
     2581 atataaatat aaagtagtaa aaattgaacc attaggagta gcacccacca aggcaaagag
     2641 aagagtggtg cagagagaaa aaagagcagt gggaatagga gctttgttcc ttgggttctt
     2701 gggagcagca ggaagcacta tgggcgcagc gtcaatgacg ctgacggtac aggccagaca
     2761 attattgtct ggtatagtgc agcagcagaa caatttgctg agggctattg aggcgcaaca
     2821 gcatctgttg caactcacag tctggggcat caagcagctc caggcaagaa tcctggctgt
     2881 ggaaagatac ctaaaggatc aacagctcct ggggatttgg ggttgctctg gaaaactcat
     2941 ttgcaccact gctgtgcctt ggaatgctag ttggagtaat aaatctctgg aacagatttg
     3001 gaatcacacg acctggatgg agtgggacag agaaattaac aattacacaa gcttaataca
     3061 ctccttaatt gaagaatcgc aaaaccagca agaaaagaat gaacaagaat tattggaatt
     3121 agataaatgg gcaagtttgt ggaattggtt taacataaca aattggctgt ggtatataaa
     3181 attattcata atgatagtag gaggcttggt aggtttaaga atagtttttg ctgtactttc
     3241 tatagtgaat agagttaggc agggatattc accattatcg tttcagaccc acctcccaac
     3301 cccgagggga cccgacaggc ccgaaggaat agaagaagaa ggtggagaga gagacagaga
     3361 cagatccatt cgattagtga acggatctcg acggtatcgc cgaattaatt cacaaatggc
     3421 agtattcatc cacaatttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     3481 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     3541 aattcaaaat tttcgggttt attacaggga cagcagagat ccagtttgga ctagtggagt
     3601 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     3661 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     3721 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     3781 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     3841 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     3901 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
     3961 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     4021 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     4081 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     4141 gacgccatcc acgctgtttt gacctccata gaagacaccg gcggccgcag atctctcgag
     4201 gttgatctgc caccatggac tacaaggatg acgatgacaa gctcgatgga ggatacccat
     4261 acgatgttcc agattacgct gctcgaggtt atcaaacaag tttgtacaaa aaagttggca
     4321 tggcggcctc attggtgggg aagaagatcg tgtttgtaac ggggaacgcc aagaagctgg
     4381 aggaggtcgt tcagattcta ggagataagt ttccatgcac tttggtggca cagaaaattg
     4441 acctgccgga gtaccagggg gagccggatg agatttccat acagaaatgt caggaggcag
     4501 ttcgccaggt acaggggccc gtgctggttg aggacacttg tctgtgcttc aatgcccttg
     4561 gagggctccc cggcccctac ataaagtggt ttctggagaa gttaaagcct gaaggtctcc
     4621 accagctcct ggccgggttc gaggacaagt cagcctatgc gctctgcacg tttgcactca
     4681 gcaccgggga cccaagccag cccgtgcgcc tgttcagggg ccggacctcg ggccggatcg
     4741 tggcacccag aggctgccag gactttggct gggacccctg ctttcagcct gatggatatg
     4801 agcagacgta cgcagagatg cctaaggcgg agaagaacgc tgtctcccat cgcttccggg
     4861 ccctgctgga gctgcaggag tactttggca gtttggcagc ttgcccaact ttcttgtaca
     4921 aagtggttcg ataacgaatt ccgccccccc cccctaacgt tactggccga agccgcttgg
     4981 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     5041 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     5101 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     5161 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     5221 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     5281 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     5341 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     5401 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     5461 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     5521 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     5581 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     5641 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     5701 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     5761 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     5821 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     5881 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     5941 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     6001 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     6061 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     6121 tgcatcgatg gatcctaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     6181 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     6241 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     6301 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     6361 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     6421 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6481 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     6541 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     6601 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     6661 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     6721 cccgcctgag atcctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     6781 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     6841 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     6901 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     6961 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7021 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7081 aaatgaatat cagagagtga gaggcccggg ttaattaagg aaagggctag atcattcttg
     7141 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     7201 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     7261 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     7321 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     7381 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     7441 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     7501 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     7561 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     7621 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     7681 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     7741 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     7801 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     7861 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     7921 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     7981 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     8041 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     8101 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     8161 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     8221 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     8281 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     8341 gtcagacccc gtagaaaaga tcaaaggatc ttc
//
