LOCUS       Exported                6170 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 6170)
  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..6170
                     /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"
     misc_feature    150
                     /label=MluI
                     /note="MluI"
     primer_bind     151..183
                     /label=HKO035
                     /note="HKO035"
     misc_feature    151..166
                     /label=pHK025-F3
                     /note="pHK025-F3"
     promoter        164..404
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    331..371
                     /label=HKO157
                     /note="HKO157"
     misc_feature    354..395
                     /label=array_F
                     /note="array_F"
     misc_feature    413..431
                     /label=chimeric RNA backbone 1 (SapI)
                     /note="chimeric RNA backbone 1 (SapI)"
     misc_feature    414..420
                     /label=SapI
                     /note="SapI"
     misc_feature    423..429
                     /label=SapI
                     /note="SapI"
     misc_RNA        448..466
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    491..515
                     /label=Readout_Rms2_priming
                     /note="Readout_Rms2_priming"
     misc_RNA        518..536
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    540..559
                     /label=Readout_BC_R_New
                     /note="Readout_BC_R_New"
     misc_feature    567..572
                     /label=U6 terminator
     promoter        726..1904
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     misc_feature    750..766
                     /label=EF1a-SR1
                     /note="EF1a-SR1"
     intron          957..1895
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     regulatory      1911..1920
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1920..2306
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     CDS             2361..2381
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2397..2939
                     /codon_start=1
                     /product="C-terminal portion of the p65 subunit of mouse 
                     NF-kappa-B"
                     /label=p65
                     /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP
                     GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE
                     FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED
                     FSSIADMDFSALLSQISS"
     CDS             2964..3335
                     /codon_start=1
                     /product="C-terminal activation domain from the human heat 
                     shock transcription factor HSF1"
                     /label=HSF1
                     /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR
                     PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED
                     PTISLLTGSEPPKAKDPTVS"
     polyA_signal    3357..3405
                     /note="synthetic polyadenylation signal"
     repeat_region   3433..3573
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3648..4103
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4385..4489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4490..5350
                     /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      5498..6165
                     /direction=RIGHT
                     /label=pUC ori
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttctag aaagagggcc tatttcccat
      181 gattccttca tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt
      241 gactgtaaac acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg
      301 gtagtttgca gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg
      361 aaagtatttc gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccggaagagc
      421 gagctcttct gttttagagc taggccaaca tgaggatcac ccatgtctgc agggcctagc
      481 aagttaaaat aaggctagtc cgttatcaac ttggccaaca tgaggatcac ccatgtctgc
      541 agggccaagt ggcaccgagt cggtgctttt ttggatccaa gcttggcgta actagatctt
      601 gagacaaatg ggacagcaga gatccagttt ggttaattag ctagctgcaa agatggataa
      661 agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg
      721 gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt
      781 tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg
      841 aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa
      901 gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa
      961 gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt
     1021 gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
     1081 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
     1141 cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
     1201 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
     1261 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
     1321 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
     1381 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
     1441 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
     1501 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
     1561 gaaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
     1621 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
     1681 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     1741 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     1801 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     1861 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgacgtacg gccaccatgg
     1921 cttcaaactt tactcagttc gtgctcgtgg acaatggtgg gacaggggat gtgacagtgg
     1981 ctccttctaa tttcgctaat ggggtggcag agtggatcag ctccaactca cggagccagg
     2041 cctacaaggt gacatgcagc gtcaggcagt ctagtgccca gaagagaaag tataccatca
     2101 aggtggaggt ccccaaagtg gctacccaga cagtgggcgg agtcgaactg cctgtcgccg
     2161 cttggaggtc ctacctgaac atggagctca ctatcccaat tttcgctacc aattctgact
     2221 gtgaactcat cgtgaaggca atgcaggggc tcctcaaaga cggtaatcct atcccttccg
     2281 ccatcgccgc taactcaggt atctacagcg ctggaggagg tggaagcgga ggaggaggaa
     2341 gcggaggagg aggtagcgga cctaagaaaa agaggaaggt ggcggccgct ggatcccctt
     2401 cagggcagat cagcaaccag gccctggctc tggcccctag ctccgctcca gtgctggccc
     2461 agactatggt gccctctagt gctatggtgc ctctggccca gccacctgct ccagcccctg
     2521 tgctgacccc aggaccaccc cagtcactga gcgctccagt gcccaagtct acacaggccg
     2581 gcgaggggac tctgagtgaa gctctgctgc acctgcagtt cgacgctgat gaggacctgg
     2641 gagctctgct ggggaacagc accgatcccg gagtgttcac agatctggcc tccgtggaca
     2701 actctgagtt tcagcagctg ctgaatcagg gcgtgtccat gtctcatagt acagccgaac
     2761 caatgctgat ggagtacccc gaagccatta cccggctggt gaccggcagc cagcggcccc
     2821 ccgaccccgc tccaactccc ctgggaacca gcggcctgcc taatgggctg tccggagatg
     2881 aagacttctc aagcatcgct gatatggact ttagtgccct gctgtcacag atttcctcta
     2941 gtgggcaggg aggaggtgga agcggcttca gcgtggacac cagtgccctg ctggacctgt
     3001 tcagcccctc ggtgaccgtg cccgacatga gcctgcctga ccttgacagc agcctggcca
     3061 gtatccaaga gctcctgtct ccccaggagc cccccaggcc tcccgaggca gagaacagca
     3121 gcccggattc agggaagcag ctggtgcact acacagcgca gccgctgttc ctgctggacc
     3181 ccggctccgt ggacaccggg agcaacgacc tgccggtgct gtttgagctg ggagagggct
     3241 cctacttctc cgaaggggac ggcttcgccg aggaccccac catctccctg ctgacaggct
     3301 cggagcctcc caaagccaag gaccccactg tctcctaaga attcgatatc aagcttaata
     3361 aaagatcttt attttcatta gatctgtgtg ttggtttttt gtgtggtaac cacgtgcgga
     3421 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     3481 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     3541 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     3601 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     3661 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3721 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3781 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3841 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3901 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3961 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     4021 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4081 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     4141 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4201 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4261 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     4321 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     4381 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     4441 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     4501 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     4561 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     4621 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     4681 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     4741 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     4801 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     4861 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4921 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4981 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     5041 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     5101 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     5161 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     5221 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     5281 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     5341 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     5401 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5461 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5521 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5581 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5641 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5701 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5761 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5821 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5881 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5941 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6001 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6061 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6121 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
