LOCUS       Exported                7084 bp ds-DNA     circular SYN 07-MAR-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7084)
  AUTHORS   John Naughton
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Mar 8, 2019 from SnapGene 4.3.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7084
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    7..136
                     /label=AAV ITR
                     /note="color: #ffcc99"
     promoter        442..719
                     /label=chicken beta-actin promoter
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    complement(1136..1169)
                     /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."
                     /note="color: #31849b; direction: LEFT"
     protein_bind    complement(1232..1265)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: LEFT"
     primer_bind     1272..1288
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     CDS             complement(1317..2036)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 1317 .. 2030 / #05fd14
                        2: 2031 .. 2033 / #05fd14 / 1a
                        3: 2034 .. 2036 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1328..1347)
                     /label=GFP Eton
                     /note="color: black; sequence: CACTCTCGGCATGGACGAGC; added:
                     2019-03-06"
     primer_bind     1886..1905
                     /label=GFP Reverse (Eton)
                     /note="color: black; sequence: TGGTGCAGATGAACTTCAGG; added:
                     2019-03-06"
     regulatory      2033..2042
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     misc_feature    complement(2037..2681)
                     /label=DTR
                     /note="color: #0000ff; direction: LEFT"
     primer_bind     complement(2707..2723)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2750..2783
                     /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."
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    2854..2887
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    2926..3514
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     primer_bind     complement(3078..3102)
                     /label=Rev07
                     /note="color: black; sequence: CTCCTCATAAAGAGACAGCAACCAG; 
                     added: 2019-03-06"
     primer_bind     3278..3297
                     /label=For08
                     /note="color: black; sequence: CTCGGCTGTTGGGCACTGAC; added:
                     2019-03-06"
     CDS             complement(3397..3408)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 3396"
                     /translation="IEGR"
     primer_bind     complement(3482..3501)
                     /label=Rev08
                     /note="color: black; sequence: CCAAAGGGAGATCCGACTCG; added:
                     2019-03-06; 5' phosphorylated"
     polyA_signal    3547..3771
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     misc_feature    3857..3986
                     /label=AAV ITR
                     /note="color: #ffcc99"
     promoter        complement(3992..4321)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      4006..4141
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     primer_bind     complement(4236..4245)
                     /label=Rev07
                     /note="color: black; sequence: CTCCTCATAAAGAGACAGCAACCAG; 
                     added: 2019-03-06"
     rep_origin      complement(4567..5155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5326..6186)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5326 .. 6117 / #ccffcc
                        2: 6118 .. 6186 / #ccffcc / signal sequence
                     Cleavage site after base 6117"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6187..6291)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 ttctagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctaggt accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      241 ctttccattg acgtcaatgg gtggactatt tacggtaaac tgcccacttg gcagtacatc
      301 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      361 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      421 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      481 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      541 tgggggcggg gggggggggg gggcgcgcgc caggcggggc ggggcggggc gaggggcggg
      601 gcggggcgag gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc
      661 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg
      721 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc
      781 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc
      841 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa
      901 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ctgtccgcgg
      961 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1021 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1081 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattgga tccccataac
     1141 ttcgtataaa gtatcctata cgaagttata tcaaaatagg aagaccaatg cttcaccatc
     1201 gacccgaatt gccaagcatc accatcgacc cataacttcg tataatgtat gctatacgaa
     1261 gttatgaatt tcgaggtcga cggtatcgat aagcttgata tcgaattccg gccgctttac
     1321 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     1381 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg
     1441 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     1501 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc
     1561 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     1621 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     1681 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     1741 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     1801 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg
     1861 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag
     1921 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     1981 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggtg
     2041 gcgaccggtg gatccgggtg ggaattagtc atgcccaact tcactttctc ttcgttttcc
     2101 acatcataac cacctctcct atggtaccta aacatgagaa gccccacgat gaccagcaga
     2161 cagacagagg acagcaccac ggccaccaca gccaggatag ttgtatggtc ataggtatat
     2221 aagcgatttt ccactgggag gctcagccca tgacacctct ctccatggta acctgggtgg
     2281 cagatgcagg agggagcccg gagctccttc acatatttgc attctccgtg gatgcagaag
     2341 tccttgtatt tccgaagaca tgggtccctc ttcttcccta gtcccttgcc tttcttcttt
     2401 cttttcccgt gctcctcctt gcttggtgtg gccagtgctt gtggcttgga ggataaagtg
     2461 actctcaaaa ggtccagatc tgcctcttgc aagtcacgga ctttccggtc ccggccgcct
     2521 cctaggcgta gcagctggtc cgtagatcca gtggaagggt ccgggttgct ggttccagca
     2581 gctagccctc tccgaagctg ctccaggctc tcgccagtca ccagtgccga aagaactgca
     2641 gccagaagga gcttcagcac caccgacggc agcagcttca tggtggcgaa ttcctgcagc
     2701 ccgggggatc cactagttct agagcggccg cctcgagaga tctgatatca taacttcgta
     2761 taggatactt tatacgaagt tatcattggg attcttccta ttttgatcca agcatcacca
     2821 tcgaccctct agtccagatc tcaccatcga cccataactt cgtatagcat acattatacg
     2881 aagttatgtc cctcgaagag gttcaagctt taacagctta tcgataatca acctctggat
     2941 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3001 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3061 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3121 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3181 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3241 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3301 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3361 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3421 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3481 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgact cgctgatcag
     3541 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     3601 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     3661 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     3721 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     3781 cggaaagaac cagctggggc tcgactagag catggctacg tagataagta gcatggcggg
     3841 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     3901 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     3961 gcctcagtga gcgagcgagc gcgcagagct ttttgcaaaa gcctaggcct ccaaaaaagc
     4021 ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa
     4081 aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga
     4141 tgggcggagt taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac
     4201 ttctgcctgc tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca
     4261 tgctttgcat acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca
     4321 cattccacag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4381 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4441 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4501 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4561 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4621 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4681 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4741 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4801 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4861 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4921 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4981 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5041 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5101 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5161 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5221 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5281 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5341 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5401 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5461 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5521 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5581 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5641 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5701 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5761 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5821 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5881 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5941 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6001 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6061 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6121 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6181 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6241 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6301 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6361 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6421 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6481 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6541 atcagagcag attgtactga gagtgcacca ttcgacgctc tcccttatgc gactcctgca
     6601 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6661 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6721 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6781 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6841 cgtagaggat ctggctagcg atgaccctgc tgattggttc gctgaccatt tccgggtgcg
     6901 ggacggcgtt accagaaact cagaaggttc gtccaaccaa accgactctg acggcagttt
     6961 acgagagaga tgatagggtc tgcttcagta agccagatgc tacacaatta ggcttgtaca
     7021 tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat
     7081 ttag
//
