LOCUS       Exported                5300 bp DNA     circular SYN 01-DEC-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5300)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2022 from SnapGene 6.0.5
            https://www.snapgene.com
COMMENT     LOCUS       pAAV-EF1a-EYFP-W        6067 bp    DNA     circular     
            20-MAY-2010

            SOURCE      

              ORGANISM  

            COMMENT     VNTDATE|538328968|

            COMMENT     VNTDBDATE|538763081|

            COMMENT     VNTNAME|pAAV-EF1a-EYFP-WPRE|

            COMMENT     VNTAUTHORNAME|William Hendriks|

            COMMENT     VNTAUTHOREML|hendriks.william@mgh.harvard.edu|

            COMMENT     VNTAUTHORNAME|Feng Zhang|

            COMMENT     VNTAUTHOREML|zhang72@fas.harvard.edu|
FEATURES             Location/Qualifiers
     source          1..5300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     gene            1..141
                     /label=L-ITR
     misc_feature    150..155
                     /label=MluI
                     /note="MluI"
     promoter        156..396
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(386..405)
                     /label=CGGTGTTTCGTCCTTTCCAC
     misc_feature    397..430
                     /label=pCM102-103_F3
                     /note="pCM102-103_F3"
     misc_RNA        425..500
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    480..519
                     /label=pCM102-103_F1
                     /note="pCM102-103_F1"
     promoter        514..754
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    729..763
                     /label=Readout_priming_site_long_F
                     /note="Readout_priming_site_long_F"
     primer_bind     complement(744..763)
                     /label=CGGTGTTTCGTCCTTTCCAC
     misc_feature    764..770
                     /label=SapI
                     /note="SapI"
     misc_feature    773..779
                     /label=SapI
                     /note="SapI"
     misc_RNA        781..856
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        925..1334
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     regulatory      1353..1362
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1389..2438
                     /label=Cre
     primer_bind     1392..1416
                     /label=CCCAAGAAGAAGAGGAAGGTGTCCA
     CDS             1392..1412
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2439..2465
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2439..2465
                     /label=HA
     polyA_signal    2487..2535
                     /note="synthetic polyadenylation signal"
     repeat_region   2563..2703
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      2778..3233
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3515..3619
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3620..4480
                     /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"
     exon            3620..4477
                     /label=AmpR
     rep_origin      4628..5295
                     /direction=RIGHT
                     /label=pUC ori
     rep_origin      4651..5239
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgaggg cctatttccc atgattcctt
      181 catatttgca tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa
      241 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
      301 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
      361 tcgatttctt ggctttatat atcttgtgga aaggacgaaa caccgtgtaa tagctcctgc
      421 atgggtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      481 aagtggcacc gagtcggtgc tttttttcta gaagagggcc tatttcccat gattccttca
      541 tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac
      601 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
      661 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
      721 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggaagagc gagctcttct
      781 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      841 ggcaccgagt cggtgctttt ttggtaccgc ggcctctaga ctcgaggggc tggaagctac
      901 ctttgacatc atttcctctg cgaatgcatg tataatttct acagaaccta ttagaaagga
      961 tcacccagcc tctgcttttg tacaactttc ccttaaaaaa ctgccaattc cactgctgtt
     1021 tggcccaata gtgagaactt tttcctgctg cctcttggtg cttttgccta tggcccctat
     1081 tctgcctgct gaagacactc ttgccagcat ggacttaaac ccctccagct ctgacaatcc
     1141 tctttctctt ttgttttaca tgaagggtct ggcagccaaa gcaatcactc aaagttcaaa
     1201 ccttatcatt ttttgctttg ttcctcttgg ccttggtttt gtacatcagc tttgaaaata
     1261 ccatcccagg gttaatgctg gggttaattt ataactaaga gtgctctagt tttgcaatac
     1321 aggacatgct ataaaaatgg aaagataccg gtgccaccat ggccccaaag gttaaccgta
     1381 cggccaccat gcccaagaag aagaggaagg tgtccaatct cctgactgtt caccagaacc
     1441 tccctgcgct gccagtagat gccactagcg atgaggtcag gaaaaatctc atggatatgt
     1501 ttagggatag acaggcgttt tctgaacaca cctggaaaat gctgcttagc gtgtgccgat
     1561 cctgggcagc ctggtgtaag ctgaacaatc gcaaatggtt ccccgccgag ccggaggacg
     1621 tgcgcgatta cctgctgtat ctccaggcaa gagggctggc tgtcaagact atccagcagc
     1681 acttgggcca actgaatatg ctgcatcgac gcagcgggct cccccggcct agcgattcaa
     1741 acgcagtctc ccttgttatg aggagaatta gaaaggaaaa cgtagatgcg ggtgagaggg
     1801 ctaagcaggc tctcgctttt gagcggactg atttcgacca ggtcagatcc ctgatggaga
     1861 acagcgatcg gtgccaggac atcaggaacc tcgcatttct gggaattgca tataacacac
     1921 ttctgcgcat agctgagatc gcccggatca gagtgaaaga catcagtcga acggacggcg
     1981 gccggatgct tattcatatt ggacgcacaa agacattggt cagcaccgct ggcgttgaaa
     2041 aggccttgtc cctgggcgta acgaagctgg tggaaagatg gatctcagtg tccggcgtgg
     2101 ctgacgaccc taataattac ttgttctgtc gagtgagaaa aaacggagtc gccgcgccct
     2161 ctgccaccag ccaattgagt acacgggccc ttgaagggat ctttgaggca acccaccgac
     2221 tcatatacgg agccaaggat gacagtggcc agaggtatct cgcctggtca ggtcattctg
     2281 ctagggtggg ggccgcacga gacatggcgc gggcaggagt ctccatacca gagattatgc
     2341 aagctggagg ttggacaaat gtgaacatcg ttatgaacta tatccgcaat cttgactctg
     2401 aaaccggggc catggtgaga ctgctcgaag atggtgacta cccatacgat gttccagatt
     2461 acgcttaaga attcgatatc aagcttaata aaagatcttt attttcatta gatctgtgtg
     2521 ttggtttttt gtgtggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     2581 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     2641 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     2701 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2761 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2821 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2881 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2941 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     3001 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3061 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     3121 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3181 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     3241 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     3301 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3361 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3421 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     3481 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     3541 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     3601 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     3661 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     3721 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     3781 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     3841 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     3901 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     3961 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4021 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4081 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4141 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4201 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     4261 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     4321 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     4381 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     4441 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     4501 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     4561 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     4621 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     4681 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     4741 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     4801 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     4861 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     4921 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     4981 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5041 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5101 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5161 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     5221 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     5281 ctggcctttt gctcacatgt
//
