LOCUS       Exported                5121 bp ds-DNA     circular SYN 08-JAN-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pVC 51 pAAV CMV mCherry NLS WPRE pA
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5121)
  AUTHORS   Engel Lab
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 8, 2021 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5121
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     protein_bind    228..249
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        264..294
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     326..342
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    379..509
                     /note="L-ITR"
     promoter        570..773
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    717..1665
                     /note="sequence verified"
     misc_feature    796
                     /note="MCS"
                     /note="multiple cloning site"
     misc_feature    802..1509
                     /label=mCherry
                     /note="mCherry"
     CDS             802..1509
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /note="mCherry"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     CDS             1522..1548
                     /codon_start=1
                     /product="nuclear localization signal of human c-myc (Dang 
                     and Lee, 1988)"
                     /note="c-myc NLS"
                     /translation="PAAKRVKLD"
     CDS             1549..1575
                     /codon_start=1
                     /product="nuclear localization signal of human c-myc (Dang 
                     and Lee, 1988)"
                     /note="c-myc NLS"
                     /translation="PAAKRVKLD"
     CDS             1576..1602
                     /codon_start=1
                     /product="nuclear localization signal of human c-myc (Dang 
                     and Lee, 1988)"
                     /note="c-myc NLS"
                     /translation="PAAKRVKLD"
     misc_feature    1630..2218
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(2101..2112)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     primer_bind     complement(2221..2237)
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    2267..2388
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     misc_feature    2550..2680
                     /note="R-ITR"
     primer_bind     complement(2698..2714)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2856..3311
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3337..3441
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             3442..4302
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4473..5061
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatctg tgatgcggtt ttggcagtac atcaatgggc
      601 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      661 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      721 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      781 tgaaccgtca gatccgctag catggtgagc aagggcgagg aggacaacat ggccatcatc
      841 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
      901 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      961 accaagggcg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
     1021 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
     1081 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
     1141 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
     1201 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
     1261 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
     1321 gacggcggcc actacgacgc cgaggtcaag accacctaca aggccaagaa gcccgtgcag
     1381 ctgcccggcg cctacaacgt caacatcaag ctggacatca cctcccacaa cgaggactac
     1441 accatcgtgg aacagtacga gcgcgccgag ggccgccact ccaccggcgg catggacgag
     1501 ctgtacaagc ctcgagctga tcctgctgcc aaacgcgtta aactagaccc tgctgccaaa
     1561 cgcgttaaac tagaccctgc tgccaaacgc gttaaactag actaaagcgg ccgcgacaag
     1621 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1681 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1741 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     1801 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     1861 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     1921 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1981 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     2041 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     2101 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2161 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     2221 cgataccgtc gacctcgacc cgggcggccg cttcgagcag acatgataag atacattgat
     2281 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2341 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2401 tgcattcatt ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa
     2461 aacctctaca aatgtggtaa aatcgataag gatcttccta gagcatggct acgtagataa
     2521 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     2581 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     2641 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc cttaattaac ctaattcact
     2701 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2761 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2821 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     2881 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2941 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3001 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3061 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3121 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3181 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3241 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3301 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     3361 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     3421 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     3481 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     3541 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3601 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3661 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3721 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3781 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3841 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3901 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3961 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     4021 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4081 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4141 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4201 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4261 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4321 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     4381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5101 tgctggcctt ttgctcacat g
//
