LOCUS       Exported                4948 bp DNA     circular SYN 24-FEB-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4948)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 30, 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..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     gene            1..141
                     /label=L-ITR
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    150..155
                     /label=MluI
                     /note="MluI"
     promoter        162..411
                     /label=Human U6 Promoter
     misc_feature    162..187
                     /label=Untitled
     promoter        163..403
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    330..370
                     /label=HKO157
                     /note="HKO157"
     misc_feature    378..412
                     /label=Readout_priming_site_long_F
                     /note="Readout_priming_site_long_F"
     misc_feature    389..412
                     /label=Readout_priming_site_F
                     /note="Readout_priming_site_F"
     misc_feature    393..411
                     /label=Untitled
     misc_feature    412..511
                     /label=chimeric RNA backbone 1
                     /note="chimeric RNA backbone 1 (SapI)"
     misc_feature    413..419
                     /label=SapI
                     /note="SapI"
     misc_feature    422..428
                     /label=SapI
                     /note="SapI"
     misc_RNA        430..505
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    430..432
                     /label=3'
                     /note="3'"
     promoter        573..982
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     regulatory      1001..1010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1040..2086
                     /label=Cre
     primer_bind     1040..1064
                     /label=CCCAAGAAGAAGAGGAAGGTGTCCA
     CDS             1040..1060
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2087..2113
                     /label=HA
     CDS             2087..2113
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2096..2125)
                     /label=ATCGAATTCTTAAGCGTAATCTGGAACATC
     polyA_signal    2135..2183
                     /note="synthetic polyadenylation signal"
     misc_feature    2135..2182
                     /label=shortPA
                     /note="shortPA"
     repeat_region   2211..2351
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      2426..2881
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3163..3267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3268..4128
                     /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"
     rep_origin      4276..4943
                     /direction=RIGHT
                     /label=pUC ori
     rep_origin      4299..4887
                     /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 cgcgttctag aagagggcct atttcccatg
      181 attccttcat atttgcatat acgatacaag gctgttagag agataattgg aattaatttg
      241 actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata atttcttggg
      301 tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac cgtaacttga
      361 aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac cggaagagcg
      421 agctcttctg ttttagagct agaaatagca agttaaaata aggctagtcc gttatcaact
      481 tgaaaaagtg gcaccgagtc ggtgcttttt tggtaccgcg gcctctagac tcgaggggtg
      541 gaagctacct ttgacatcat ttcctctgcg aatgcatgta taatttctac agaacctatt
      601 agaaaggatc acccagcctc tgcttttgta caactttccc ttaaaaaact gccaattcca
      661 ctgctgtttg gcccaatagt gagaactttt tcctgctgcc tcttggtgct tttgcctatg
      721 gcccctattc tgcctgctga agacactctt gccagcatgg acttaaaccc ctccagctct
      781 gacaatcctc tttctctttt gttttacatg aagggtctgg cagccaaagc aatcactcaa
      841 agttcaaacc ttatcatttt ttgctttgtt cctcttggcc ttggttttgt acatcagctt
      901 tgaaaatacc atcccagggt taatgctggg gttaatttat aactaagagt gctctagttt
      961 tgcaatacag gacatgctat aaaaatggaa agataccggt gccaccatgg ccccaaaggt
     1021 taaccgtacg gccaccatgc ccaagaagaa gaggaaggtg tccaatctcc tgactgttca
     1081 ccagaacctc cctgcgctgc cagtagatgc cactagcgat gaggtcagga aaaatctcat
     1141 ggatatgttt agggatagac aggcgttttc tgaacacacc tggaaaatgc tgcttagcgt
     1201 gtgccgatcc tgggcagcct ggtgtaagct gaacaatcgc aaatggttcc ccgccgagcc
     1261 ggaggacgtg cgcgattacc tgctgtatct ccaggcaaga gggctggctg tcaagactat
     1321 ccagcagcac ttgggccaac tgaatatgct gcatcgacgc agcgggctcc cccggcctag
     1381 cgattcaaac gcagtctccc ttgttatgag gagaattaga aaggaaaacg tagatgcggg
     1441 tgagagggct aagcaggctc tcgcttttga gcggactgat ttcgaccagg tcagatccct
     1501 gatggagaac agcgatcggt gccaggacat caggaacctc gcatttctgg gaattgcata
     1561 taacacactt ctgcgcatag ctgagatcgc ccggatcaga gtgaaagaca tcagtcgaac
     1621 ggacggcggc cggatgctta ttcatattgg acgcacaaag acattggtca gcaccgctgg
     1681 cgttgaaaag gccttgtccc tgggcgtaac gaagctggtg gaaagatgga tctcagtgtc
     1741 cggcgtggct gacgacccta ataattactt gttctgtcga gtgagaaaaa acggagtcgc
     1801 cgcgccctct gccaccagcc aattgagtac acgggccctt gaagggatct ttgaggcaac
     1861 ccaccgactc atatacggag ccaaggatga cagtggccag aggtatctcg cctggtcagg
     1921 tcattctgct agggtggggg ccgcacgaga catggcgcgg gcaggagtct ccataccaga
     1981 gattatgcaa gctggaggtt ggacaaatgt gaacatcgtt atgaactata tccgcaatct
     2041 tgactctgaa accggggcca tggtgagact gctcgaagat ggtgactacc catacgatgt
     2101 tccagattac gcttaagaat tcgatatcaa gcttaataaa agatctttat tttcattaga
     2161 tctgtgtgtt ggttttttgt gtggtaacca cgtgcggacc gagcggccgc aggaacccct
     2221 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     2281 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     2341 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     2401 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2461 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2521 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2581 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2641 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2701 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2761 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2821 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     2881 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     2941 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3001 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3061 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     3121 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3181 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3241 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3301 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3361 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3421 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3481 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     3541 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3601 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3661 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3721 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3781 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3841 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3901 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3961 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4021 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4081 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4141 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4201 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4261 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4321 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4381 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4441 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4501 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4561 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4621 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4681 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4741 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4801 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4861 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4921 gccttttgct ggccttttgc tcacatgt
//
