pAAVS1-EF1a-puro-DNR

LOCUS       pAAVS1-EF1a-puro-DNR        8077 bp DNA     circular SYN 26-OCT-2024
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8077)
  AUTHORS   Ming
  TITLE     Direct Submission
  JOURNAL   Exported Oct 26, 2024 from SnapGene 6.0.2 to Vector NTI(R) format
            https://www.biofeng.com
COMMENT     VNTDATE|1072224000|
COMMENT     VNTDBDATE|1072224000|
COMMENT     VNTNAME|pAAVS1-EF1a-puro-DNR|
COMMENT     VNTAUTHORNAME|Ming|
FEATURES             Location/Qualifiers
     primer_bind     complement(95..111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    119..135
                     /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)."
     promoter        complement(143..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    188..209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(1256..2116)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     promoter        complement(2117..2221)
                     /gene="bla"
                     /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"
     CDS             complement(4381..4980)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     promoter        complement(5020..5519)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        5580..6758
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          5811..6749
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     CDS             6873..6902
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
     CDS             6921..6944
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
     primer_bind     complement(6964..6980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    6991..7467
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     misc_feature    7542..8071
                     /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 ttgcggggcg 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 tcagcgtgag
     5581 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     5641 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     5701 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     5761 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     5821 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     5881 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     5941 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     6001 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     6061 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     6121 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     6181 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     6241 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     6301 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     6361 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     6421 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     6481 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     6541 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     6601 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     6661 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     6721 tggttcaaag tttttttctt ccatttcagg tgtcgtgagc ggccgggaat tcgtcgactg
     6781 gatccggtac cgaggagatc tgccgccgcg atcgccggcg cgccagatct caagcttaac
     6841 tagctagcgg accgacgcgt acgcggccgc tcgagcagaa actcatctca gaagaggatc
     6901 tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa acggccggcc
     6961 gcggtcatag ctgtttcctg aacagatccc gggtggcatc cctgtgaccc ctccccagtg
     7021 cctctcctgg ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta
     7081 agttgcatca ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg
     7141 gtatggagca aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa
     7201 ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa
     7261 gcgattctcc tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc
     7321 agctaatttt tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc
     7381 aactcctaat ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg
     7441 tgaaccactg ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac
     7501 gtccagacac agcataggct acctggccat gcccaaccgg tgccagagag gatcctggga
     7561 gggagagctt ggcagggggt gggagggaag ggggggatgc gtgacctgcc cggttctcag
     7621 tggccaccct gcgctaccct ctcccagaac ctgagctgct ctgacgcggc cgtctggtgc
     7681 gtttcactga tcctggtgct gcagcttcct tacacttccc aagaggagaa gcagtttgga
     7741 aaaacaaaat cagaataagt tggtcctgag ttctaacttt ggctcttcac ctttctagtc
     7801 cccaatttat attgttcctc cgtgcgtcag ttttacctgt gagataaggc cagtagccag
     7861 ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg tgtggaaaac tccctttgtg
     7921 agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc tctactccct ttctctttct
     7981 ccatccttct ttccttaaag agtccccagt gctatctggg acatattcct ccgcccagag
     8041 cagggtcccg cttccctaag gccctgctct gtctaga
//