AAVS1-EF1a

LOCUS       AAVS1-EF1a              6991 bp ds-DNA     circular SYN 04-MAY-2016
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6991)
  AUTHORS   Youbio
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Sep 28, 2022 from SnapGene Viewer 5.1.4.1 to 
            Vector NTI(R) format
            https://www.biofeng.com
COMMENT     VNTDATE|768960000|
COMMENT     VNTDBDATE|768960000|
COMMENT     VNTNAME|AAVS1-EF1a|
COMMENT     VNTAUTHORNAME|Youbio|
FEATURES             Location/Qualifiers
     misc_feature    33..836
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    843..868
                     /label=SA
                     /note="splice acceptor site"
     CDS             892..945
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
     CDS             955..1554
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
     polyA_signal    1592..1816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1822..2201
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2308..2519
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2532..2800
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     polyA_site      3030..3085
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     misc_feature    3634..4470
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     CDS             complement(4976..5836)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      5960..6548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        6971..6989
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagtcctgc aggtttaaac gaattcgccc tttgctttct ctgaccagca ttctctcccc
       61 tgggcctgtg ccgctttctg tctgcagctt gtggcctggg tcacctctac ggctggccca
      121 gatccttccc tgccgcctcc ttcaggttcc gtcttcctcc actccctctt ccccttgctc
      181 tctgctgtgt tgctgcccaa ggatgctctt tccggagcac ttccttctcg gcgctgcacc
      241 acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc ctctccgggc atctctcctc
      301 cctcacccaa ccccatgccg tcttcactcg ctgggttccc ttttccttct ccttctgggg
      361 cctgtgccat ctctcgtttc ttaggatggc cttctccgac ggatgtctcc cttgcgtccc
      421 gcctcccctt cttgtaggcc tgcatcatca ccgtttttct ggacaacccc aaagtacccc
      481 gtctccctgg ctttagccac ctctccatcc tcttgctttc tttgcctgga caccccgttc
      541 tcctgtggat tcgggtcacc tctcactcct ttcatttggg cagctcccct acccccctta
      601 cctctctagt ctgtgctagc tcttccagcc ccctgtcatg gcatcttcca ggggtccgag
      661 agctcagcta gtcttcttcc tccaacccgg gcccctatgt ccacttcagg acagcatgtt
      721 tgctgcctcc agggatcctg tgtccccgag ctgggaccac cttatattcc cagggccggt
      781 taatgtggct ctggttctgg gtacttttat ctgtcccctc caccccacag tggggcaagc
      841 ttctgacctc ttctcttcct cccacagggc ctcgagagat ctggcagcgg agagggcaga
      901 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgagatgacc
      961 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     1021 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     1081 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     1141 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     1201 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     1261 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     1321 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     1381 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     1441 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     1501 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatctaga
     1561 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1621 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1681 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1741 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1801 gcggtgggct ctatgggtct cgacattgat tattgactag ttattaatag taatcaatta
     1861 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1921 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1981 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2041 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2101 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2161 cttggcagta catctacgta ttagtcatcg ctattaccat ggaatagcaa cagacataca
     2221 aactaaagaa ttacaaaaac aaattacaaa aattcaaaat tttatcgata ctagtaagga
     2281 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     2341 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact
     2401 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     2461 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc
     2521 tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc
     2581 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt
     2641 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt
     2701 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct
     2761 acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta
     2821 ctctagagct agcgaattcg aaggacttaa gggtaccgat atcaccggtg tcgacacgcg
     2881 tgaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     2941 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     3001 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     3061 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     3121 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     3181 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3241 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3301 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     3361 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct
     3421 gcagcccaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3481 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3541 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3601 tgtcgtgcca gcggatcgac agtactaagc tttactaggg acaggattgg tgacagaaaa
     3661 gccccatcct taggcctcct ccttcctagt ctcctgatat tgggtctaac ccccacctcc
     3721 tgttaggcag attccttatc tggtgacaca cccccatttc ctggagccat ctctctcctt
     3781 gccagaacct ctaaggtttg cttacgatgg agccagagag gatcctggga gggagagctt
     3841 ggcagggggt gggagggaag ggggggatgc gtgacctgcc cggttctcag tggccaccct
     3901 gcgctaccct ctcccagaac ctgagctgct ctgacgcggc tgtctggtgc gtttcactga
     3961 tcctggtgct gcagcttcct tacacttccc aagaggagaa gcagtttgga aaaacaaaat
     4021 cagaataagt tggtcctgag ttctaacttt ggctcttcac ctttctagtc cccaatttat
     4081 attgttcctc cgtgcgtcag ttttacctgt gagataaggc cagtagccag ccccgtcctg
     4141 gcagggctgt ggtgaggagg ggggtgtccg tgtggaaaac tccctttgtg agaatggtgc
     4201 gtcctaggtg ttcaccaggt cgtggccgcc tctactccct ttctctttct ccatccttct
     4261 ttccttaaag agtccccagt gctatctggg acatattcct ccgcccagag cagggtcccg
     4321 cttccctaag gccctgctct gggcttctgg gtttgagtcc ttggcaagcc caggagaggc
     4381 gctcaggctt ccctgtcccc cttcctcgtc caccatctca tgcccctggc tctcctgccc
     4441 cttccctaca ggggttcctg gctctgctct aagggcaagg gcgaattcgc ggcctggcga
     4501 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4561 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4621 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4681 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgaatta ttaacgctta
     4741 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatcag
     4801 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4861 caaatatgta tccgctcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     4921 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     4981 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5041 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5101 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5161 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5221 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5281 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5341 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5401 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5461 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5521 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5581 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5641 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5701 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     5761 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     5821 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     5881 gtctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5941 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6001 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6061 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6121 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6181 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6241 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6301 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6361 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6421 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6481 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6541 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6601 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6661 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6721 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6781 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6841 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6901 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     6961 ccaagctcag aattaaccct cactaaaggg a
//