LOCUS       Exported                8956 bp ds-DNA     circular SYN 22-JUN-2021
DEFINITION  synthetic circular DNA
ACCESSION   urn.local...14-bo81j1k
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8956)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 2, 2021 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     primer_bind     4..20
                     /label=M13 rev
     misc_feature    complement(61..196)
                     /label=PiggyBac 5'TR
     misc_feature    77..95
                     /label=TR2
     primer_bind     complement(356..379)
                     /label=Rv2 - PiggyBac NO attP NO lox EM835
     misc_feature    complement(371..591)
                     /label=attP'
     protein_bind    427..526
                     /label=phage phi-C31 attP
     protein_bind    602..635
                     /label=loxP
     misc_feature    638..640
                     /label=lac promoter
     promoter        645..2560
                     /label=Nix promoter
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    645..668
                     /label=Nix Fwd4
                     /note="/created_by=User"
     exon            2570..3315
                     /label=Exon 1
                     /note="/modified_by=User"
     misc_feature    2570..2641
                     /label=5' UTR
     misc_feature    2642..2644
                     /label=Start
     misc_feature    complement(3290..3315)
                     /label=Aal Nix E1 Rvs
     exon            3316..3380
                     /label=Exon 3
                     /note="/modified_by=User"
     misc_feature    3316..3334
                     /label=Aal Nix E3 Fwd
     misc_feature    complement(3355..3380)
                     /label=Aal Nix E3 Rvs
     exon            3381..3722
                     /label=Exon 4
                     /note="/modified_by=User"
     misc_feature    3381..3404
                     /label=Aal Nix E4 Fwd
     misc_feature    3399..3401
                     /label=agg
     misc_feature    3481..3483
                     /label=stop?
     misc_feature    3482..3722
                     /label=3' UTR
     misc_feature    complement(3699..3723)
                     /label=Aal Nix E4 Rvs
     primer_bind     3724..3750
                     /label=EM820
     misc_feature    3728..3950
                     /label=SV40 term
     polyA_signal    3831..3913
                     /label=SV40 poly(A) signal
     promoter        3959..5339
                     /label=PUb
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    5347..6063
                     /label=YFP
                     /note="/created_by=User"
     misc_feature    6109..6346
                     /label=SV40
     misc_feature    6228..6346
                     /label=SV40 polyA signal
     primer_bind     complement(6325..6346)
                     /label=EM1121
     protein_bind    6364..6397
                     /label=loxP
     misc_feature    6411..6506
                     /label=repeated transposase region
     misc_feature    complement(6490..6647)
                     /label=original PiggyBac 3'region
     misc_feature    complement(6585..6603)
                     /label=TR2
     misc_feature    6599..6619
                     /label=direct repeat
     misc_feature    complement(6635..6647)
                     /label=TR1
     primer_bind     complement(6761..6810)
                     /label=EM854
     primer_bind     complement(6777..6793)
                     /label=M13 fwd
     misc_feature    6795..6826
                     /label=BbsI sites GGGG-CCAT
     primer_bind     6811..6846
                     /label=EM841
     misc_feature    join(6825..8968,1..33)
                     /label=pDONR backbone
     terminator      6958..6985
                     /label=rrnB T2 terminator
     primer_bind     complement(7216..7237)
                     /label=EM599
     rep_origin      complement(7315..7903)
                     /direction=LEFT
                     /label=ori
     CDS             complement(8050..8859)
                     /label=KanR
     misc_feature    8930..8948
                     /label=pDONR-RP
     misc_feature    6062..6102
                     /note="SV40 NLS (no effect)"
     source          1..8968
                     /dnas_title="pB_Nix_OE_E1E3E4_YFPcorrecetd"
ORIGIN      
        1 tgccaggaaa cagctatgac catgtaatac gacgatatga tcctgatgca gctagattaa
       61 ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt
      121 tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc
      181 ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt
      241 gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact catacgataa
      301 ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt
      361 atagattaga tcgcgctcgc gcgactgacg gtcgtaagca cccgcgtacg tgtccacccc
      421 ggtcacaacc ccttgtgtca tgtcggcgac cctacgcccc caactgagag aactcaaagg
      481 ttaccccagt tggggcacta ctcccgaaaa ccgcttctga cctgggaaaa cgtgaagccc
      541 cggggcatcc gctgagggtt gccgccgggg cttcggtgtg tccgtcagta ctgcaggtac
      601 cataacttcg tatagcatac attatacgaa gttataccgg atcctcgcat tttatgagta
      661 aaggcccatt actcatatat gggtaaagtg ctttttgtaa aaaatgagta aatcgattta
      721 ctcataaatt ataatttcac cgtgtttact tttcgtcttt tgttatattt ttgaactgtt
      781 aactattttg tatttatcat ttaattcata caaaattttg cgtttatttt gtagtttatt
      841 gcaatctacg aaaattatta atgaattttt acatgtttca aatactctta atcattttcc
      901 tcctggaaaa cagaccttcc gagacgtagc tctttgaagt tttcataatg atccatgtat
      961 gaaattttcc cctccggctg ttgttcatcg cgcagttgtt cctgttcttc cgttcctcgt
     1021 tcttccggcg cttgaagcct tggtgcagtc ggaactgtgg aaTaagactg atatattttt
     1081 atttcacggt cgttaatgaa aatgatgctg tcactatgga acttacctgc aaattccatg
     1141 cttttgtagc ctttcttgta gctccacatg atgatttgtt gctcctgtat tatcttcgag
     1201 taccaaacct tgtgaaacgg aaataatcca tcgtaaatga aaattgaacg gtatcaattt
     1261 atccaggaag tacttaccga ttaaaatatc aatgtatttg ggaacctcga ctctaatccg
     1321 cgtgggaatc cgatggacgc ggtatcggtc attgaacccg atgctgaagg gcctcgactg
     1381 gctatagcgg tgccgtgaaa atcaaaatat ctaaaatttc ccgatagtgt tatgatcagg
     1441 ggatccgcat gcggcaaggt aggcctactg ctcatctttt gcgagtgtgt tagtgtgact
     1501 ggcaacgcga ccagaagtct agtcgaatcc tgcttggaaa tcggtttgtt ttgtcgatgg
     1561 gggtggcccg cgaaatcgca ttgtgttgtt tgggaaggcc cgctcgatgc atgttaggta
     1621 ggcgtagttt gttcgagttt gacgtgcctg caccctccct ggttatgatg gcgcactaat
     1681 gcattccgag aaaagttgct aacttgaaaa ttttcatcca aatatgggta agttcattta
     1741 cccatgtttg ggtatagcga ttttacctat aatatgggca aatttgaccg tttttcaaag
     1801 gtatatttta cccatattat gggtaatgca tatgaaaccc aaatatgggt aaatcaacta
     1861 ctgatctatg ggtaaactga tcttagcgtg tatgtttcac acgtgacatt cactggtttt
     1921 acaattggct tctttagcag tgttgggata attccatatt atgaatctgc atgccaaact
     1981 gggccgaaat ccaaattttc atcaattttg gtgcacggga acctatttaa atatcaattt
     2041 gaagtttgta tgggagcgat ttgtcgaatc acccctcgtt gcattttgta ctgggcggag
     2101 ctgtcaaaca gttgcctagc tgtcaaaagg tgatttcgaa taatctcttt gaaattgatt
     2161 ttaggtatca aaataaagtt ctaaaaatct gaaaaaaatc atagtggctc agaaaaaggt
     2221 gctctttcgt ataaaatcaa aaaatgaaca cttttttcaa aatttaaaaa cccaattttc
     2281 gcaaatctat agggctctgc acgaagttct ctccctctct ttcgctctca ttgagatttt
     2341 gtaaacaaca aggccaggaa atgtcaaaat cccatacaaa atcaaaacag tgcagtgccc
     2401 tatatgtaaa acacatcact ccgacgtgta aatttttttg agtgttgatt taatcaaagt
     2461 gaataaaaat attagtttta tgacatactt gttttctgag tgtagcaaaa atatgaaaac
     2521 acatttttgt actttgaatg ttaagcgtgt atgctttttg GTTTGGTTAt gtcaatatgt
     2581 caattgttaa acccatgtta atagttttaa ttttttttta atcaaattct ttttttaagt
     2641 aatgtacagt aaaagtgaac ttaatctcat taacaatcaa tttgaataca ttaaaaaata
     2701 ttgcatatac attggaaaca ttcccgccga agtatcgaaa acagatttaa ttgcaaaatt
     2761 ttccgtattt ggtgaaatat ctaacttata catgaagtca ttcattcagt tttgtgatgt
     2821 gaaaccggca gttgttcgtt acagactgat gaaaagtgta aaggaatctt caagtttaca
     2881 caatagtcga tatattcaat cggttttaat agttctgcca ctagattctt cctacaataa
     2941 ttactttctt ccttacaaca cttgtgttgt ggtatacact tataacaaat ttggcatggt
     3001 agatttttat caaaaattca gtaaattagg agatatacat gcgatgaaga aagctacaaa
     3061 tgtcatggtt tacattagct ttgtatcaga aagagctgca aggaccattc tggatactaa
     3121 gcctacagat atacatataa atgtacaaac aattaatcat gttacacgaa atattaacgt
     3181 atgcttaata gattttgaaa aggaatgtac atcaaatacg gcgataaaat taacactttt
     3241 atataatcgc tcaattggaa tattcggact accatctaat ttcacagaag caaaactgca
     3301 cgatgaattt tcaagctgcg gaccgttcaa gaaattttgg cagcggaaac atttcttgac
     3361 cttttccaca atacttatca gctttggaaa atccccagag gtagatcaat cctactccgc
     3421 caaaaacttc cctggcgatg gaagctcttc cgaaccgatc ttcccgagtc ggtgtcgagc
     3481 tagattctgt tttttccggt tggagactgA tctccggttc actggcgccc cagcgaccgt
     3541 gctttccatg acgttatcga tctagcctac tctcccggcg gtggacctag cctactctga
     3601 tggaaaaaat ccccagcgaa agaagttctc ccgaaaccga gaattcaatc tcgaccatcc
     3661 aggtgccgag gtcgacccag cgatgccggt tggaggttga cttccagttc actatcactc
     3721 cgataagcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3781 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3841 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3901 ttttcttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc taagAGTTat
     3961 ctttacatgt agcttgtgca ttgaatccaa ttataatttg ccttggcacc agctgagcca
     4021 gacaagaaag aaagcttccc agaagtatat cgatttagaa gggttgacgt cactttgctg
     4081 actgcactaa tacagcaaat gatacaatta gaatgattca agtgaaattc ccaaattact
     4141 gctttgtctc tggatttggt tatcagatta cattcgaagc taagaatagc taccgaaatt
     4201 gtcgatcaaa tcaggaaatc ctttctctat cgaaaaaggc attcgcacat ctttctctgt
     4261 atgccatata cacgaatggt aggtacattg acgtctttgc cagaagttga actgcatcgt
     4321 tcaaggtaca gaatgaacga ctaacagaca caagcacgtt ttgctgtcca ttcagacaca
     4381 gggatggtac ccatagtcga tcgatttaga gccatccaac cgaacagagg tatatgtatg
     4441 aatggattgc agaaattttc tagaagtaca accaccacta cggcagtgtc tataaaacgc
     4501 ccctgcaaag gcaaaaccag ctcaatcgaa tacgtttcct agtggagtga acattacgcg
     4561 gtccaagtaa gcagtgccag tgcaagtgaa gtgaagtctc tagtgaaaaa gagtgatcca
     4621 attagccaga ggagaaaatt tcagagtgaa caaagctttg ttcaaaggac aattactatt
     4681 aaatttgtga aagtgcattt cggtgaaggg aatcttctag tgaaggtagg taaattaaat
     4741 gatgaaatta tagctatgag cgaaaactag tttggtgaat gattcctttg tctttgaatg
     4801 agcaaactat tttccaagat ggcgactatt gagctttgag tgattagtga aaatttgcaa
     4861 cgcagtttca tcatcattga taaaacccaa ttgtgattca cggcgataat catatttcgt
     4921 tgaatcatcg ctgctaattg aattaaattt ctagagcaag cgcgaattcg ccatatttct
     4981 aaaattaaat attgtggtga taattaccca ttaaggtaat attaacacat atcgagaaaa
     5041 accttgagga aatcgtgaaa acttgaagat acgcaatttc caaactacgt agttcaaagt
     5101 cgaaaacaag ttaatttttc actaaaaagt agggcgttgt tgtgacgtca tcaccttcaa
     5161 gtgtatattt ttcacttggc ctgcgactgc aaacgcagac aaagcaaaac aagtttaaaa
     5221 cctgtcgtgt cgtgctcgaa gccaaaggca atgaatcaat atcaaatgag agtttgcatt
     5281 tcacaaccaa ttactgaagc gtttcctcgt ttctttttct gctcaacaga gatttcaacG
     5341 TTCACCatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     5401 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     5461 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     5521 cccaccctcg tgaccacctt cggctacggc ctgcagtgct tcgcccgcta ccccgaccac
     5581 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     5641 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     5701 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     5761 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     5821 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     5881 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     5941 aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     6001 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     6061 aagaGATCTC GACCCAAGAA AAAGCGGAAG GTGGAGGACC CGTAAagcgg ccgcgactct
     6121 agatcataat cagccatacc acatttgtag aggttTTact tgctttaaaa aacctcccac
     6181 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     6241 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     6301 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcgctt gtaattcgtc
     6361 gacataactt cgtatagcat acattatacg aagttatgag ctcaattcga taaaagtttt
     6421 gttactttat agaagaaatt ttgagttttt gtttttttta ataaataaat aaacataaat
     6481 aaattgtttg ttgaatttat tattagtatg taagtgtaaa tataataaaa cttaatatct
     6541 attcaaatta ataaataaac ctcgatatac agaccgataa aacacatgcg tcaattttac
     6601 gcatgattat ctttaacgta cgtcacaata tgattatctt tctagggtta atctagctgc
     6661 gtgttctgca gcgtgtcgag catcttcatc tgctccatca cgctgtaaaa cacatttgca
     6721 ccgcgagtct gcccgtcctc cacgggttca aaaacgtgaa tgaacgaggc gcgctcactg
     6781 gccgtcgttt tacaggggat gtcttcatat atatgaagac tcccatctgt tgtttgtcgg
     6841 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gtgaagcaac
     6901 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa actaagcaga
     6961 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttcctggc tagcggtacg
     7021 cgtattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     7081 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     7141 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     7201 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     7261 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     7321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     7381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     7441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     7501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7741 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7921 ttctacgggg tctgacgctc agtggaacga cgcgtaactc acgttaaggg attttggtca
     7981 tgagcttgcg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac
     8041 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     8101 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     8161 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     8221 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     8281 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     8341 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     8401 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     8461 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     8521 atcaggatat tcttctaata cctggaatgc tgtttttccg gggatcgcag tggtgagtaa
     8581 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     8641 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     8701 tttcagaaac aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga
     8761 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     8821 taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     8881 tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca
     8941 tcagagattt tgagacacgg gccagagc
//

