LOCUS       pAAVS1-Puro-DNR.        7507 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Mammalian donor vector for transgene integration at the AAVS1 locus 
            in the human genome. Use together with pCas-Guide-AAVS1.
ACCESSION   .
VERSION     .
KEYWORDS    pAAVS1-Puro-DNR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7507)
  AUTHORS   OriGene
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7507)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(119..135)
                     /label=lac operator
                     /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)."
     promoter        complement(143..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(188..209)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(497..1085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1259..2116)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2117..2221)
                     /label=AmpR promoter
     rep_origin      complement(2503..2958)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3171..3187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3278..3779
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     protein_bind    complement(3792..3824)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    complement(3851..4304)
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation
                     signal"
     CDS             complement(4384..4980)
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     promoter        complement(5020..5519)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     enhancer        5637..5940
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5941..6144
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        6170..6188
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    6197..6300
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             6303..6332
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             6351..6374
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     primer_bind     complement(6394..6410)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6421..6897
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     misc_feature    6972..7501
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
ORIGIN
        1 atcactctcg ccggttggac tttagatcag aagggatctt gctgccgccc gaaagaggaa
       61 gggctggaag aggaaggagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
      121 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
      181 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
      241 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
      301 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
      361 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      421 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      481 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      541 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      601 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      661 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      721 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      781 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      841 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      901 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      961 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1021 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1081 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1141 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     1201 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1261 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1321 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1381 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     1441 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1501 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1561 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1621 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1681 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1741 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1801 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1861 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1921 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1981 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     2041 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     2101 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     2161 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     2221 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     2281 cctataaaaa taggcgtatc acgaggccct ttcgggtcgc gcgtttcggt gatgacggtg
     2341 aaaacctctg acacatgcag ctcccgttga cggtcacagc ttgtctgtaa gcggatgccg
     2401 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     2461 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
     2521 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
     2581 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
     2641 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2701 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
     2761 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2821 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2881 aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2941 cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag
     3001 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     3061 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     3121 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     3181 ggccagtgaa ttggaggcta cagtcagtgg agaggacttt cacaggctgt cgccgtgctc
     3241 atttgataac tgcccgttat tcatgcgaca ctctagagag cacttccttc tcggcgctgc
     3301 accacgtgat gtcctctgag cggatcctcc ccgtgtctgg gtcctctccg ggcatctctc
     3361 ctccctcacc caaccccatg ccgtcttcac tcgctgggtt cccttttcct tctccttctg
     3421 gggcctgtgc catctctcgt ttcttaggat ggccttctcc gacggatgtc tcccttgcgt
     3481 cccgcctccc cttcttgtag gcctgcatca tcaccgtttt tctggacaac cccaaagtac
     3541 cccgtctccc tggctttagc cacctctcca tcctcttgct ttctttgcct ggacaccccg
     3601 ttctcctgtg gattcgggtc acctctcact cctttcattt gggcagctcc cctacccccc
     3661 ttacctctct agtctgtgct agctcttcca gccccctgtc atggcatctt ccaggggtcc
     3721 gagagctcag ctagtcttct tcctccaacc cgggccccta tgtccacttc aggacagcac
     3781 tagtggacct aataacttct atagcataca ttatacgaag ttatattaag ggttccggat
     3841 ctcgaccagc ttctgatgga attagaactt ggcaaaacaa tactgagaat gaagtgtatg
     3901 tggaacagag gctgctgatc tcgttcttca ggctatgaaa ctgacacatt tggaaaccac
     3961 agtacttaga accacaaagt gggaatcaag agaaaaacaa tgatcccacg agagatctat
     4021 agatctatag atcatgagtg ggaggaatga gctggccctt aatttggttt tgcttgttta
     4081 aattatgata tccaactatg aaacattatc ataaagcaat agtaaagagc cttcagtaaa
     4141 gagcaggcat ttatctaatc ccaccccacc cccacccccg tagctccaat ccttccattc
     4201 aaaatgtagg tactctgttc tcacccttct taacaaagta tgacaggaaa aacttccatt
     4261 ttagtggaca tctttattgt ttaatagatc atcaatttct gcatcccggg gatctgatat
     4321 catcgatgca tggggtcgtg cgctcctttc ggtcgggcgc tgcgggtcgt ggggcgggcg
     4381 tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc
     4441 ggcggtgacg gtgaagccga gccgctcgta gaaggggagg ttgtggggcg cggaggtctc
     4501 caggaaggcg ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct
     4561 gcccagaccc ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg
     4621 ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga
     4681 accgctcaac tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct
     4741 ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat
     4801 gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg
     4861 gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg
     4921 tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat
     4981 ggtaagcttc agctgctcga gatctagatg gatgcaggtc gaaaggcccg gagatgagga
     5041 agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg ggcctccgga
     5101 ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg agcccgcccc cggacccacc
     5161 ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc tgctattggc cgctgcccca
     5221 aaggcctacc cgcttccatt gctcagcggt gctgtccatc tgcacgagac tggtgagacg
     5281 tgctacttcc atttgtcacg tcctgcacga cgcgagctgc ggggcggggg ggaacttcct
     5341 gactagggga ggagtagaag gtggcgcgaa ggggccacca aagaacggag tcggttggcg
     5401 cctaccggtg gatgtggaat gtgtgcgagg ccagaggcca cttgtgtagc gccaagtgcc
     5461 cagcggggct gctaaagcgc atgctccaga ctgccttggg aaaagcgcct cccctacccg
     5521 gtagaatttc gaggtcgaga tcctaagctt ggctggacgt aaactcctct tcagactagt
     5581 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     5641 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     5701 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     5761 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     5821 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     5881 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     5941 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
     6001 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     6061 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
     6121 tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattttgt aatacgactc
     6181 actatagggc ggccgggaat tcgtcgactg gatccggtac cgaggagatc tgccgccgcg
     6241 atcgccggcg cgccagatct caagcttaac tagctagcgg accgacgcgt acgcggccgc
     6301 tcgagcagaa actcatctca gaagaggatc tggcagcaaa tgatatcctg gattacaagg
     6361 atgacgacga taaggtttaa acggccggcc gcggtcatag ctgtttcctg aacagatccc
     6421 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     6481 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     6541 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     6601 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     6661 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     6721 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     6781 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     6841 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     6901 attttaaaat aactatacca gcaggaggac gtccagacac agcataggct acctggccat
     6961 gcccaaccgg tgccagagag gatcctggga gggagagctt ggcagggggt gggagggaag
     7021 ggggggatgc gtgacctgcc cggttctcag tggccaccct gcgctaccct ctcccagaac
     7081 ctgagctgct ctgacgcggc cgtctggtgc gtttcactga tcctggtgct gcagcttcct
     7141 tacacttccc aagaggagaa gcagtttgga aaaacaaaat cagaataagt tggtcctgag
     7201 ttctaacttt ggctcttcac ctttctagtc cccaatttat attgttcctc cgtgcgtcag
     7261 ttttacctgt gagataaggc cagtagccag ccccgtcctg gcagggctgt ggtgaggagg
     7321 ggggtgtccg tgtggaaaac tccctttgtg agaatggtgc gtcctaggtg ttcaccaggt
     7381 cgtggccgcc tctactccct ttctctttct ccatccttct ttccttaaag agtccccagt
     7441 gctatctggg acatattcct ccgcccagag cagggtcccg cttccctaag gccctgctct
     7501 gtctaga
//
