LOCUS       Exported                7073 bp ds-DNA     circular SYN 06-MAR-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7073)
  AUTHORS   kevinu
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Oct 25, 2019 from SnapGene Viewer 5.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7073
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    164..648
                     /label=hSyn promoter
     protein_bind    complement(680..713)
                     /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."
     protein_bind    complement(776..809)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(830..1549)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(1550..1606)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    complement(1634..3004)
                     /label=Tetanus toxin light chain
     protein_bind    3021..3054
                     /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."
     primer_bind     complement(3061..3083)
                     /label=BT_Seq_F
     protein_bind    3125..3158
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3200..3788
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(3671..3682)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3820..4296
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   4336..4476
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4551..5006
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5288..5392
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5393..6253
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEELFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDYTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPHGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6424..7012
                     /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 cgcgtttaat taagtgtcta gactgcagag
      181 ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct
      241 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc
      301 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag
      361 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca
      421 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc
      481 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag
      541 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg
      601 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgagaaac cggctagagg
      661 atccgccgcc accatggcca taacttcgta taaagtatcc tatacgaagt tatatcaaaa
      721 taggaagacc aatgcttcac catcgacccg aattgccaag catcaccatc gacccataac
      781 ttcgtataat gtatgctata cgaagttatt tcgcctgcag gggcgcgcct tacttgtaca
      841 gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat
      901 cgcgcttctc gttggggtct ttgctcaggg cggactggta gctcaggtag tggttgtcgg
      961 gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca
     1021 cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct
     1081 tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga
     1141 tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc
     1201 cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc
     1261 gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt
     1321 cggggtagcg ggcgaagcac tgcaggccgt agccgaaggt ggtcacgagg gtgggccagg
     1381 gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat
     1441 cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga
     1501 ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatt ggcccgggat
     1561 tctcttcgac atcccctgct tgtttcaaca gggagaagtt agtggctccg cttccgttaa
     1621 cagcgctgct agcagcggta cggttgtaca ggttttcacg gatgttggtc ggcgggatga
     1681 tttttttgca cagaccgatc agtttggaaa ccagaccgga accgtcgacg ttcctaaagg
     1741 cgttagtgtt caccctcatg ttctggccct tatactcgga cttcagatcc ttggactcta
     1801 tgttaaagcc ctcagtatcg ttatatatag tatcatccag caggttaggt atcttcacag
     1861 gatcgtggtt catggaaaaa tagcttaagc gagtctttat gttaaacttc ttgcccagct
     1921 ctatctcagt aaagccatac attatggagt tatacagtat ctgaaactta tcctcgttca
     1981 ctatatactg gccgttggaa tccttatcaa actgatactt ctgctgatag atctgtttgt
     2041 aggagtcgat gtcgatgttc gggtcgttgc aggaggtaac ctgggacagt ttgttagcga
     2101 tagctttgta gtcgttcagg gttttttcgt acaggtcgtt tttgatatcg atggagatca
     2161 ggttagcgtc ctgaccaccg aaggtgaaga gctcttcagc ggagatcggg taggtgtgct
     2221 gcatgtagat ttcctgtttg gacgggatga tttcgtggga ggaaacctgc atgccgtaca
     2281 gaccgtgcag aacgtggatc agttcgtgca tcagcagcag agccgggtcc tggaagtatt
     2341 tggatttacc gatggtcagg gaggtgatgt tttcgataac gttgtcgaag gtcggtacgt
     2401 attccgggca gaaagccatc tgcatgatgg aaccgaaacc gtcacggcac gggaagtagt
     2461 ttttgttgtc aacacgcaga acgataccac gaacttcgtt tttgttcaga accggacccg
     2521 gaccgaagat gatcaggttg gtcagcatag cggatttggt ggtagcaccg gacggatcct
     2581 gttccagcag gttgaaggaa acggagttgg agttggtgtc gaatttgtcc agcagggagt
     2641 aggagttacc caggtacggg atagcgttga tgattttgtc cagcagagct tcaccagcaa
     2701 cgttgttttt gatacggttg aacagtttaa ccatggtctg caggaaacgg tctttgtcgg
     2761 agtcggtacg caggtagttc gggtcgtagt attcggaagc accttcgatc agggaggacg
     2821 gcgggttgaa gtcttccggt ttggtaccaa attcgtaacg ttccggaacg atccagatac
     2881 ggtcggtgat tttgaaggct ttgtagtaga tgtctagacc tttgcagtac ggcggttcca
     2941 tcatgatgat ggtgtcgttg ttaaccgggt cggagtaacg gaagttgttg atggtgatcg
     3001 gcataccggt ttctcgacgc ataacttcgt ataggatact ttatacgaag ttatctttgg
     3061 gattcttcct attttgatcc aagcatcacc atcgagggtc tagtccagat ctcaccatcg
     3121 acccataact tcgtatagca tacattatac gaagttatgt ccctcgaaga ggttcgaatt
     3181 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3241 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3301 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3361 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3421 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3481 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3541 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3601 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     3661 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3721 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3781 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     3841 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3901 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3961 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     4021 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     4081 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     4141 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     4201 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     4261 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
     4321 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4381 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4441 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4501 ttacgcatct gtgcggtatt tcacaccgca tacgtgaaag caaccatagt acgcgccctg
     4561 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4621 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4681 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4741 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4801 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4861 ccaaactgga acaacactca accctatctc gggctattct tttgatttat aagggatttt
     4921 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4981 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     5041 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     5101 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     5161 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     5221 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     5281 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     5341 catgagagaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5401 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5461 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5521 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaact
     5581 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5641 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5701 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5761 tgccataacc atgagtgatt acactgcggc caacttactt ctgacaacga tcggaggacc
     5821 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5881 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5941 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     6001 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     6061 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     6121 cattgcagca ctggggccac atggtaagcc ctcccgtatc gtagttatct acacgacggg
     6181 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     6241 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     6301 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6361 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6421 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     6481 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     6541 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     6601 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     6661 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     6721 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     6781 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     6841 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     6901 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     6961 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7021 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
