LOCUS       Exported                6298 bp ds-DNA     circular SYN 01-MAR-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6298)
  AUTHORS   Paul Sabatini
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 28, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6298
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    298..568
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        601..638
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    612..632
                     /label=283
     protein_bind    773..806
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    817..850
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     CDS             complement(906..1616)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(986..1009)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     1083..1099
                     /label=ps254
                     /note="hM3Dq R2 (mcitrine)"
     primer_bind     complement(1184..1203)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1293..1303
                     /label=ps254
                     /note="hM3Dq R2 (mcitrine)"
     primer_bind     1453..1471
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1599..1618)
                     /label=ps121
                     /note="dsGFP F AscI"
     primer_bind     complement(1599..1616)
                     /label=ps117
                     /note="dsGFP F MluI"
     primer_bind     complement(1599..1616)
                     /label=ps119
                     /note="dsGFP F AgeI"
     regulatory      1613..1622
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(1807..1826)
                     /label=ps197
                     /note="Coff/Fon ChR2 R seq validation"
     primer_bind     1993..2011
                     /label=ps198
                     /note="Con/Fon ChR2 R seq validation (use with 196)"
     primer_bind     2075..2095
                     /label=ps261
                     /note="Ai27 ChR2 R"
     primer_bind     complement(2508..2527)
                     /label=ps260
                     /note="Ai27 ChR2 F"
     regulatory      2555..2564
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(2601..2634)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(2645..2678)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    2702..3290
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2755..2775)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3173..3184)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3325..3342)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3331..3538
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3561..3701
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3561..3690
                     /label=AAV2 ITR
     rep_origin      3776..4231
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3863..3882)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4073..4094
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4248..4267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4367..4389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4427..4445)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4513..4617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4618..5478
                     /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"
     primer_bind     complement(4836..4855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5649..6237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6138..6157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgaaacaa ttctgcagga attcggggcc
      181 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
      241 ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggtcgagt ttaccactcc
      301 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      361 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      421 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      481 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      541 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga
      601 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccgcggccc cgaattcgag ctcggtaccc ggggatcctc tagtcgacgt cgataacttc
      781 gtatagcata cattatacga agttatccga atcgcaataa cttcgtatag gatactttat
      841 acgaagttat ttggcgcgcc aaatttaaat aacgcaacgt accttcgagg gggcccaagg
      901 aattcttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgctcg
      961 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccagctt gatgttgacg
     1021 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcggcg
     1081 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     1141 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     1201 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     1261 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     1321 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     1381 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     1441 agggggccgc ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     1501 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     1561 aagcgcatga actccttgat gatggccatg ttgtcctcct cgcccttgct caccatggtg
     1621 gcgaccggtg ccacggctcc ggcctcggct tcgtcttcga cgagagtctc gacctcgatc
     1681 tccgttccgc cgatgttcag tttggtggtt ttgcggatat ctccgtgaat caatatgtgc
     1741 tcgtggatca ggacgcgcag gtagtgtccc aacaaccccc aacaattttt actcatcaga
     1801 tcaataatcg tgtgacctac ggtggagcca tagacgctca ggacgccaaa accttcgggc
     1861 cccaaaatga agagaattgg gaacataccc cagctcacga aaaacagcca tgccatgccg
     1921 gtcacgacct ggcggcaccg accctttggc acagtatgat aaccctcgat atatgctttg
     1981 gcggcgtgaa aaaatgtgtt cgcgccatag cacaatccaa gacaaaagaa gatgacttta
     2041 acatagccgg ttgccatggc gctggtagcc ccccacacga tagtcccgat gtctgagaca
     2101 aggagtccca tggttctcct gctgtagtcg ttgctcaggc cggtgaggtt gctcaggcgg
     2161 ataaggatga caggacaagt gagcagccac tctgcatagc gcagccactg cacccggtgt
     2221 cctgtggcaa ggtagagcat agagggattc ttaaactcaa aaaagaactc gagaatcacc
     2281 ttaaccattt caatggcgca cacatagatc tcctcccagc cgcatgtaga tttccaggtt
     2341 tggtaggcat agaacatcag cagcaaaatg ctgaatcctg ctgcaagcca ctgcaggaca
     2401 tttgacgcgg tctgagcgcc gttcgtgccg cgagattcaa tccatccggc acagtaacat
     2461 tgatcctcag ggaccaggac ggacccgttc accaccacag gattagtaac gaacaaaagt
     2521 tcgcgtccga cggcagacaa agcgccgcca tagtccatgg tggccgctag ctagttggcg
     2581 cgccaacctg gccggccttg ataacttcgt ataatgtatg ctatacgaag ttatccgaat
     2641 cgcaataact tcgtataaag tatcctatac gaagttatcg aagatcatca agcttatcga
     2701 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2761 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2821 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2881 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2941 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3001 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3061 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3121 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3181 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3241 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3301 agcgctgctc gagagatcga tctgcctcga ctgtgccttc tagttgccag ccatctgttg
     3361 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3421 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3481 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggaca
     3541 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     3601 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3661 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     3721 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     3781 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3841 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3901 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     3961 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4021 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4081 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg
     4141 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4201 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4261 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4321 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4381 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4441 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4501 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4561 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4621 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4681 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4741 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4801 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4861 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4921 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4981 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5041 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5101 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5161 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5221 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5281 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5341 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5401 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5461 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5521 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5581 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5641 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5701 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5761 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5821 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5881 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5941 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6001 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6061 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6121 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6181 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6241 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
