LOCUS       pAAV.2xSV40pA.E_        6136 bp DNA     circular SYN 29-OCT-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6136)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 1, 2023 from SnapGene 7.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      449..1037
                     /label=ori
     primer_bind     1028..1046
                     /label=pAAV.seq1
     repeat_region   1099..1239
                     /label=L-ITR
     primer_bind     1238..1265
                     /label=JG97.STOP.L-ITR_IF.for
     misc_feature    1253..1280
                     /label=3xSV40pA.FOR
     misc_feature    1301..1809
                     /label=2xSV40pA
     primer_bind     complement(1415..1437)
                     /label=Enrich_AAVGenomeBAR_P7_R
     polyA_signal    1421..1555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1500..1523)
                     /label=SH72.pAAV_seq8R
     primer_bind     complement(1669..1691)
                     /label=Enrich_AAVGenomeBAR_P7_R
     polyA_signal    1675..1809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1754..1777)
                     /label=SH72.pAAV_seq8R
     primer_bind     complement(1807..1832)
                     /label=JG99.STOP.Sst044_IF.rev
     misc_feature    complement(1810..1831)
                     /label=3xSV40pA.REV
     primer_bind     1822..1858
                     /label=JG178.Sst44.AvrII_IF.for
     misc_feature    1838..2557
                     /label=Sst44
     primer_bind     complement(2540..2572)
                     /label=JG183.Sst44.pCMV*_IF.rev
     primer_bind     2558..2579
                     /label=JG172.pCMV*.for
     misc_feature    2564..2695
                     /label=mCMV*_minP
     primer_bind     complement(2674..2723)
                     /label=JG90.mCMV*_minP.SpeI_IF.rev
     primer_bind     2709..2803
                     /label=JG182.mTagBFP2.NcoI_IF.for
     primer_bind     2709..2774
                     /label=JG227.FLAG.SpeI_IF.for
     misc_feature    2715..2720
                     /label=Kozak
     CDS             2721..2726
                     /codon_start=1
                     /label=Start
                     /translation="MV"
     CDS             2727..2747
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2754..2777
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2784..3491
                     /codon_start=1
                     /label=mTagBFP2
                     /translation="VSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDM
                     ALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVA
                     RYCDLPSKLGHKLN"
     primer_bind     complement(3470..3538)
                     /label=JG128.nlsBFP2.AscI_IF.rev
     CDS             3498..3518
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     3560..3597
                     /label=JG141.WPRE3.ERexp_IF.for
     misc_feature    3565..4153
                     /label=WPRE
     primer_bind     complement(3571..3600)
                     /label=JG81.seq4R
     primer_bind     3742..3759
                     /label=pAAV.seq5
     primer_bind     4122..4140
                     /label=pAAV.seq6
     misc_feature    4131..4186
                     /label=56_1
     misc_feature    4144..4199
                     /label=UTR2_read2
     primer_bind     4144..4160
                     /label=R2_Viral_UTR2
     primer_bind     complement(4151..4176)
                     /label=SH92
     misc_feature    4152..4207
                     /label=UTR1_read2
     primer_bind     4152..4168
                     /label=R2_Viral_UTR1
     misc_feature    4177..4232
                     /label=56_2
     misc_feature    4177..4186
                     /label=Barcode
     primer_bind     complement(4331..4353)
                     /label=Enrich_AAVGenomeBAR_P7_R
     polyA_signal    4337..4418
                     /label=SV40 poly(A) signal
     misc_feature    4368..4373
                     /label=polyA
     misc_feature    4397..4402
                     /label=polyA
     primer_bind     complement(4416..4439)
                     /label=SH72.pAAV_seq8R
     primer_bind     complement(4470..4489)
                     /label=JG50.pAAV.seq7R
     repeat_region   4497..4637
                     /label=R-ITR
     rep_origin      4712..5167
                     /label=F1 Origin
     promoter        5449..5553
                     /label=AmpR promoter
     CDS             5554..5622
                     /label=AmpR
     CDS             join(5623..6136,1..278)
                     /label=AmpR
ORIGIN
        1 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
       61 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
      121 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
      181 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
      241 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
      301 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
      361 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
      421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
      481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
      541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
      601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
      661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
      901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1081 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     1141 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     1201 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgcacg cgttcgcgat
     1261 gaataaatga aagcttgcag atctgcgact ctagaggatc tgcgactcta gaggatcata
     1321 atcagccata ccacattttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     1381 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     1441 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1501 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgcga
     1561 ctctagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     1621 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     1681 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     1741 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     1801 gtctggatcc ccatcaagct gatccggaac ccctagggtt cagtacccag acactcccat
     1861 acccttattt agaagaaata aatatcatca agtcataata tccttgactg attaagaaag
     1921 ccactttgta agtgtttatt aaactgtcaa gaaacttaca gaatttacta catgatcgtt
     1981 agaataactt tgagtcagga catatttgat atgacttaat catactccct ccaaaaggaa
     2041 ataaggcttt gtgaaggtaa attatttctt cctgggttgg atatgtgttt atggagtgat
     2101 cattcagctg ttcccaacct tcattctgaa aaggcctcag aacacttcat gatgaatcaa
     2161 gctgtatcct gaatagagta aaatgaacca cttcgtagga actatggtgt caccacatca
     2221 gcaattctta ttgaaaagtg tgcatttctt attcacatat ttcaaagatg gtattccaga
     2281 ggagtgattt tctcaatgta tttttcatct acaagccttc attttaagcc taccaccgtg
     2341 tgtgttttca agacagcaat tatcgtttta aaatgtgcag gtctagcttg agcttctcag
     2401 caagtttcta tgccaaagaa aacaccaatc ctttccattt actgagaatc aatgtttaat
     2461 cctccttttt gttctcatac ttattacaaa tcataaagaa ttctgagtgt cagtttgata
     2521 actagaagct ccatgtacca ttcctgctcc ttattgagtc gacgtataag ctttaggcgt
     2581 gtacggtggg cgcctataaa agcagagctc gtttagtgaa ccgtcagatc gcctggagca
     2641 attccacaac acttttgtct tataccaact ttccgtacca cttcctaccc tcgtaaactc
     2701 ttaagaaact agtagccacc atggtgccaa agaagaagcg taaggtagga tccgactaca
     2761 aggacgacga cgacaaagga tccgtgtcta agggcgaaga gctgattaag gagaacatgc
     2821 acatgaagct gtacatggag ggcaccgtgg acaaccatca cttcaagtgc acatccgagg
     2881 gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc gagggcggcc
     2941 ctctcccctt cgccttcgac atcctggcta ctagcttcct ctacggcagc aagaccttca
     3001 tcaaccacac ccagggcatc cccgacttct tcaagcagtc cttccctgag ggcttcacat
     3061 gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag gacaccagcc
     3121 tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc acatccaacg
     3181 gccctgtgat gcagaagaaa acactcggct gggaggcctt caccgagacg ctgtaccccg
     3241 ctgacggcgg cctggaaggc agaaacgaca tggccctgaa gctcgtgggc gggagccatc
     3301 tgatcgcaaa cgccaagacc acatatagat ccaagaaacc cgctaagaac ctcaagatgc
     3361 ctggcgtcta ctatgtggac tacagactgg aaagaatcaa ggaggccaac aacgagacct
     3421 acgtcgagca gcacgaggtg gcagtggcca gatactgcga cctccctagc aaactggggc
     3481 acaagcttaa tggaagcccc aagaaaaagc ggaaggtgta aggcgcgcca cccctgcagg
     3541 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     3601 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     3661 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     3721 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     3781 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     3841 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     3901 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     3961 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc
     4021 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4081 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4141 gccgcctccc cgcatcgata ccgagcgcgc gatcgcttcc gcataatcga gagatctgtg
     4201 atagcggcca tcaagctggc cgcgactcta gatcataatc agccatacca catttgtaga
     4261 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4321 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4381 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4441 actcatcaat gtatcagctt atcgataccg catgcacgtg cggaccgagc ggccgcagga
     4501 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4561 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4621 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4681 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4741 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4801 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4861 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4921 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4981 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5041 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5101 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5161 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5221 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5281 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5341 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5401 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5461 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5521 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5581 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5641 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5701 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5761 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5821 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5881 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5941 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6001 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6061 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6121 aaactattaa ctggcg
//
