LOCUS       Exported                7847 bp ds-DNA     circular SYN 18-JAN-2022
DEFINITION  Created in 20170321.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7847)
  AUTHORS   Bing Xu
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 19, 2022 from SnapGene 4.3.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7847
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    748..1277
                     /label=Homologous arm
                     /note="color: #a6acb3"
     enhancer        1284..1477
                     /label=Xef1 Enhancer
                     /note="color: #ff668e"
     promoter        1478..2964
                     /label=actb2 promoter
                     /note="color: #fcd74e"
     CDS             4426..4428
                     /codon_start=1
                     /label=N-mApple-ATG
                     /note="color: #ff0000"
                     /translation="M"
     misc_feature    4446..4477
                     /label=rox
                     /note="color: #008000; direction: RIGHT"
     protein_bind    4480..4513
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    4514..4547
                     /label=FRT5
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    4548..4581
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACAT). loxP511 sites are compatible with each
                     other, but incompatible with wild-type loxP sites."
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    4582..4615
                     /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    4616..4649
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     CDS             4769..5476
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="VSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEAF
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNFE
                     DGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALKS
                     EIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     polyA_signal    5507..5628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     LTR             complement(5750..6266)
                     /label=Left arm of Tol2
                     /note="vntifkey=19"
                     /note="color: #000080; direction: LEFT"
     rep_origin      complement(6794..7382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             7553..7575
                     /codon_start=1
                     /label=Ampicillin_stop
                     /note="vntifkey=4"
                     /note="color: #3366cc"
                     /translation="LPMLNQ"
     CDS             complement(7810..7847)
                     /codon_start=1
                     /label=Ampicillin
                     /note="vntifkey=4"
                     /note="color: #993366"
                     /translation="QTINWRTTYSSF"
     CDS             complement(7847)
                     /codon_start=1
                     /label=Ampicillin_start
                     /note="vntifkey=4"
                     /note="color: #993366"
                     /translation=""
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacgc tggctaagaa
      241 ctcatcagcc tccccggtcc atctacccac gtaccaatgc accaattggc cacaatgacg
      301 gctactacat ggtgccattc cttcctcttt ataggaatgg agactacctc ctgtccaaca
      361 aggctcttgg atacgagtac gcctacctgt tggacccagg tcattgcaca acaccagaaa
      421 tgccctctga tctgcaaaag acgtgaatat ctgttcagac acccatatcc actctgttcc
      481 acacaggtca gaggtttgtc caggagttct tgacagaggt gtaaaaagta ctcaaaaatt
      541 ttactcaagt gaaagtacaa gtacttaggg aaaattttac tcaattaaaa gtaaaagtat
      601 ctggctagaa tcttacttga gtaaaagtaa aaaagtactc cattaaaatt gtacttgagt
      661 attaaggaag taaaagtaaa agcaagaaag aaaactagag attcttgttt aagcttttaa
      721 tctcaaaaaa cattaaatga aatgcatttc cagatgagtt caccaccctg tgctttaacg
      781 tgactgcagg agcaacactc ttaagtctct ccccaatcag cgtatgtgaa gcgtgagagt
      841 gagagttaga agataaatga ggttttttag tttatttttg gttgctggag gttggcttga
      901 aggtgagatc ctttcgtaga caaaattact caatcaattt gtatggctgg taggttgagt
      961 tagccttgga ctgcttatgt aatggggttg aaaacttgag ccccttctgt tctgcaccat
     1021 taacatcacc atttatcacc tttagctgaa atcttcattc tcacttttca ctctctaaaa
     1081 ctgcttttgg caatgtttcg cggatcttat gttggtgttt ttcaatgtat gatcgcgttt
     1141 caggccatta aatggatttt tacttcaagt ctgatcaata ggcggcttaa ctggacccaa
     1201 gtgctttctt aggctgtcag tatctttaga ggaaggtgaa gtgatattag aaatccaaat
     1261 ttaggcctgt aaggaaactc gagcaggggg atcatctaat caagcacaaa taaggggcgt
     1321 gtaacacaaa agccagcgac cctttccaat gcaaatcaaa cttgcaattc tttgccgttt
     1381 ttatcattta agtgtcggct taaggtccac tatcagatgt aaacagcctt atctaacaaa
     1441 ggtatcatta cattctgaaa ttctcaggca tgcaagctag agtgctttta caatgtagat
     1501 tgtgtcaaag cagcttaact tagaggatta gtatagtaaa ttgaaactgt gtcagtccat
     1561 tttttaagag tttaagttca gttcagttta gttcagtgtg agttaatttt tactgctgag
     1621 agtccaaaca ctaaagagca aatccaaata cacaagtcaa taaataaact caatgatggg
     1681 ctaaagatct ggggaaatct gcagatttct gtgcgcacag attctgtgtg ggcctagtca
     1741 ttacctatac tcaacgttta ctctgagaaa cttccatatt ttagaggtaa aatgctcaga
     1801 attgtaaaaa gtcttgtcaa aatacttgga atttgccaaa aacaaaataa aagtcacatc
     1861 acctcattgt ccttggtatc agcaagcact tttaacacaa tcagctgttt ttgtttataa
     1921 ctcccactga aggccatgtg cattactccc aagttacaca gtaaaaagtc acatgtagtt
     1981 tatcatgatc actgctgctc aagcacagct ttatctgtta ttcagtgtgg caatatctga
     2041 aatgtgtgca aaatattatt tattgtagct gatcaccaac actttcagtg tttttttttt
     2101 ctacttttaa tgagaattat gttcttagga tttatcctta taaacctctt taaaaagaaa
     2161 tgttatgtct gcatagacac gacatgaaat tatgaacaga aaatttaact atttaagatc
     2221 aagacttgac atataacatt tctaatgaac attaaacctg aggtaggccc acttgcaaaa
     2281 atgtaactgc tatcactcac tttatacatc attgtgtaac ttgtacacac aaaaatagtg
     2341 aagcaatgtg gcatggaatg caggtcccat tcattcaaag ttattaaagc actttcttaa
     2401 agtgcattac tactgagcta agaccagcta gttgctcata caacatacta caagtagtgt
     2461 catcctaatt tggataaaaa aaatgtccat tttatacatg gaaaagtact cattagatcc
     2521 cccattgttt gtattacggt atttcgtgaa cacaagaggt aaatgaccta cagagctgct
     2581 gttgtgttag attgaaaaca caacacagga tcatggaggc atccatcatt atgacgcatt
     2641 ctgccacttg taactctgca cagacttttt gaaaagttta attgagtctt ggcgttgtcg
     2701 cagaaaaata cgaaaaattc tgtgttaaaa tttgcaaata aaaattccag ttttatagga
     2761 aatcgccatg gccttgttct ttaaccagtt aagcttcccc ttctttcact ctcaagttgc
     2821 aagaagcaag tgtagcaatg tgcacgcgac agccgggtgt gtgacgctgg accaatcaga
     2881 gcacagagct ccgaaagttt accttttatg gctagagccg ggtatgtgcc gtcatataaa
     2941 agagcgcgcc cagcttttca gcctcacttt gagctcctcc acacgcagct agtgcggaat
     3001 atcatctgct tgtaacccat tctcttaagt cgacaacccc ccaaacccaa ggtgagttgg
     3061 ttttacagtt tttaagcgtt taattatagt ttctatctat taatgttaag taaaatgtgt
     3121 aaatggataa atatgatttg atgaagtctt tagacagttc ataacggtct ctttgtagtt
     3181 atctactagg tttgatttat aagtggtggt tcttttgata atctatgtta catttaataa
     3241 atattggctg tatcttaact tttttgtagc gttagtggat gtggcaggtg agaatgtcgg
     3301 taatttaacg tgaccagtcg taggcacgac attgaacatt gaataggccg gtgtgaaatg
     3361 ggtgttaagt cttaatataa agttgtgttc tggctaatct ctctttaaca gccataaatg
     3421 tgatttttaa ctacagtttt gagggtgctg gtgagcatgt tgcacacttg atggatagcc
     3481 ggcatgggaa gttctttgtg caggcagtgc tgcagcaggg tgtgacctac tttagctagc
     3541 cggctaacca gctctcatct gatgtaacgt aaaccccatc aatctaacat cggctcattc
     3601 gtgctttata agatcgaaga tcttgttatt agtaacgggt attggcattt agttttaact
     3661 tggttaattt tctttttgtt atgtgttcta tttaaatgta gcctgcagcc ccctctgtct
     3721 ttaatgaagc tgaagattaa cgttaacgtt agtgaagtgt tgtatccgct atgaaagatt
     3781 tgaattttag aggtgtggtc cactttaata acggctgcgg agttctctgg ctgcacttca
     3841 gccgctgtga ctgtcgaagc agagcccatc agtttttcag gatctcggtg atttttgagg
     3901 ctagactatg aactgaaccg actgagagtg accgtccaga aatgctttag tgtgaagtct
     3961 ccgctcacga ggagtccgtg gactgactgc agatctgtga cgcagtaaca cccggcggca
     4021 gacacccgtt gaaatgcggt tgtgtaattg atcgcaggcg agggtggaag aggatgtgaa
     4081 acttcatttg tgtaaaattt agggagtggt cctggtgtga tgaatgtcga aatctgttcc
     4141 tttttactga gccctacgac tctggctgag tgccacaccg ccggcagccg caaagcggct
     4201 caaaccattg ccttttatgg taataatgag agaatgcaga gggacttcct ttgtctggca
     4261 tatctgaggc gcgcgctgtc actagcgccc accagcggtc agactgtaga atgcagagca
     4321 aaacaggaag ttgactccag atggtcacat gcttgctgaa acgcctctcc ctggcagcag
     4381 tgcaattcta acagtttccc tttcttttac agtggatccg ccaccatggt tggtatcaaa
     4441 gatcttaact ttaaataatg ccaattattt aaagttaccg aagttcctat tctctagaaa
     4501 gtataggaac ttcgaagttc ctattcttca aaaggtatag gaacttcata acttcgtata
     4561 gtacacatta tacgaagtta tataacttcg tataggatac tttatacgaa gttatataac
     4621 ttcgtatagc atacattata cgaagttatg gttacaagac aggtttaagg agaccaatag
     4681 aaactgggca tgtggagaca gagaagactc ttgggtttct gataggcact gactctctct
     4741 gcctattggt ctattttccc acccttaggt gagcaagggc gaggagaata acatggccat
     4801 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt
     4861 cgagatcgag ggcgagggcg agggccgccc ctacgaggcc tttcagaccg ctaagctgaa
     4921 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta
     4981 cggctccaag gtctacatta agcacccagc cgacatcccc gactacttca agctgtcctt
     5041 ccccgagggc ttcaggtggg agcgcgtgat gaacttcgag gacggcggca ttattcacgt
     5101 taaccaggac tcctccctgc aggacggcgt gttcatctac aaggtgaagc tgcgcggcac
     5161 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccga
     5221 ggagcggatg taccccgagg acggcgccct gaagagcgag atcaagaaga ggctgaagct
     5281 gaaggacggc ggccactacg ccgccgaggt caagaccacc tacaaggcca agaagcccgt
     5341 gcagctgccc ggcgcctaca tcgtcgacat caagttggac atcgtgtccc acaacgagga
     5401 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga
     5461 cgagctgtac aagtaacata tgtaagtata cttgatatcg acatgataag atacattgat
     5521 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     5581 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     5641 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa
     5701 aacctctaca aatgtggtat ggctgattat gatcctctag atcagatctg cgaagatacg
     5761 gccacgggtg ctcttgatcc tgtggctgat tttggactgt gctgctcgca gctgctgatg
     5821 aatcacatac ttcctccatt ttcttccact gattgactgt tataatttcc ctaatttcca
     5881 ggtcaaggtg ctgtgcattg tggtaataga tgtgacatga cgtcacttcc aaaggaccaa
     5941 tgaacatgtc tgaccaattt catataatgt gaaaacgatt ttcataggca gaataaataa
     6001 catttaaatt aaactgggca tcagcgcaat tcaattggtt tggtaatagc aagggaaaat
     6061 agaatgaagt gatctccaaa aaataagtac tttttgactg taaataaaat tgtaaggagt
     6121 aaaaagtact tttttttcta aaaaaatgta attaagtaaa agtaaaagta ttgattttta
     6181 attgtactca agtaaagtaa aaatccccaa aaataatact taagtacagt aatcaagtaa
     6241 aattactcaa gtactttaca cctctggttc ttgaccccct accttcagca agcccagcag
     6301 atccactagt tctagagcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt
     6361 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6421 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6481 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6541 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6601 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7201 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     7261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     7381 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7441 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7501 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7561 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7621 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7681 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7741 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7801 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttg
//
