LOCUS       Exported                9117 bp ds-DNA     circular SYN 21-DEC-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 9117)
  AUTHORS   CIM-Functional Validation Core
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 20, 2022 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9117
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    complement(222..413)
                     /label=Tol2_ITR-R
     misc_feature    430..447
                     /label=I-SceI
     misc_recomb     448..481
                     /label=LoxP
     misc_feature    488..597
                     /label=BP/SA
                     /note="carp beta actin splice acceptor from intron 1/exon2 
                     boundary"
     CDS             651..1325
                     /label=mRFP*
                     /note="mRFP with AUG removed"
     polyA_signal    1338..2513
                     /label=Ocean Pout Terminator
                     /note="Ocean pout antifreeze protein 3'UTR, poly(A) signal,
                     and putative boundary element (Pat Gibbs)"
     misc_feature    2602..3737
                     /label=beta-actin promoter
                     /note="Carp beta-actin promoter"
     exon            3743..3834
                     /label=beta actin exon 1
                     /note="carp beta actin exon 1 (non-coding)"
     intron          3835..5098
                     /label=beta-actin intron 1
                     /note="carp beta actin intron 1"
     CDS             5163..5879
                     /label=GFP
                     /note="GM2 variant"
     misc_feature    5943..6070
                     /label=SD
                     /note="carp beta actin splice-donor exon/intron 1 boundary"
     misc_recomb     6088..6121
                     /label=loxP
     misc_feature    6120..6137
                     /label=I-SceI
     misc_feature    6138..6399
                     /label=Tol2_ITR-L
     rep_origin      6594..7049
                     /label=F1 Ori
     CDS             complement(7232..8092)
                     /label=AmpR
     rep_origin      complement(8153..9033)
                     /direction=LEFT
                     /label=ColE1 Ori
ORIGIN
        1 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
       61 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      121 aggaaacagc tatgaccatg attacgccaa gctatttagg tgacactata gaatactcaa
      181 gctatgcatc caacgcgttg ggagctctcc catatggtcg agcagaggtg taaaaagtac
      241 tcaaaaattt tactcaagtg aaagtacaag tacttaggga aaattttact caattaaaag
      301 taaaagtatc tggctagaat cttacttgag taaaagtaaa aaagtactcc attaaaattg
      361 tacttgagta ttaaggaagt aaaagtaaaa gcaagaaaga aaactagaga ttcttgttta
      421 agctcatggt agggataaca gggtaatata acttcgtata gcatacatta tacgaagtta
      481 tcgttaccac ccactagcgg tcagactgca gattgcagca cgaaacagga agctgactcc
      541 acatggtcac atgctcactg aagtgttgac ttccctgaca gctgtgcact ttctaaaccg
      601 gttttctcat tcatttacag cagccggaat tcacccgggt accggaaggt gcctcctccg
      661 aggacgtcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc
      721 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
      781 agctgaaggt gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
      841 tccagtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
      901 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
      961 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     1021 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg
     1081 cctccaccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc
     1141 tgaagctgaa ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga
     1201 agcccgtgca gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca
     1261 acgaggacta caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg
     1321 cctaagaaat cctagtaatt aagtctcagc caccgttaac tgaacatgtc aaaacctgtg
     1381 gagactgttg agatttgatg ttctgaaaag ataaagccta taaataaaat gttgcccaaa
     1441 tttcctgcct gatgtttttc tttgtctttg ctacatggct ttgctgctcg gatcggctca
     1501 ctctgtgtat gccacgttca ctttgtactc tccttctcac ggtaggttta ttatttttag
     1561 atgtgcagtt agtttctgtg aaataacaca ccacacactg atattgtctg tgcattgact
     1621 tggtgagtgc acattgtttt tgatcttgac atatttatat ttgattgatc aggtgaactg
     1681 tgtgaatcta aagtgctcca tacagatgtt ctgcattgaa aatattctca ttttattagt
     1741 ggaagtgagt gtatgccaca tccaatcaat ttcagcaaac accccagtat gatttaatgc
     1801 aaaaaaatga aggtatcaaa cacgcattac tactttgcag ttaaatattt aacatttatt
     1861 ccaacacgaa aaaaagcagt aaataacact ttgacaaaca cgtcaggaca tcttattttt
     1921 gtcaccctca caggcaattt agtataatat attatatata tatatatatc atataataat
     1981 attcagtata atatatatat atatatcata ttataatatt cagtataata taaaacacaa
     2041 acacatatat gtataatata atataacatt tttatttatt gagatgcctc tatggaccgt
     2101 gttataagaa gtaaagatca ggagaagtaa acatgaagtg taattatgaa tactgatgtt
     2161 aaattaagct atgatgagtt ttcactgtta atttaccatc tcaattaaat gttgatgcct
     2221 ccatgaccaa gttaagcaga tgagactgag acaactgtag aagacaagat gttcactttg
     2281 ctgaatatag ctggcttgac agttatctat gactctataa atatatatat attttttttt
     2341 ttataaaatg atttatttat aactatatat ccatttctca gacaggtgct tcatatccct
     2401 cactcccgta gctgtccatg ctggatctgt ccccgttgtt tttaaaaagc taaataagtt
     2461 attaacatga ctgcatccag cgagccaaac ctgtctggtg tacagctacc agagaagctt
     2521 gagatcctag gaagttccta tactttctag agaataggaa tcgcgagtgc tagcaattaa
     2581 ccctcactaa agggaacaaa agcttttaga ccttcttact tttggggatt atataagtat
     2641 tttctcaata aatatctcat atcttactgt ggtttaactg ctgaatctaa aattttaata
     2701 caaaagtagt tatatttgtt gtacattgta aactataact taacttcagt ttcagagaaa
     2761 ctcatgtgct caaaatgtaa aaaaagtttc ctgttaaata ttttgtaaat gtattgaaga
     2821 caaaataaga aaaaaaaaaa tataagccac taaatcacac tgtccttggt atcagcaaga
     2881 gattctgaca taatcagctg tttttgttta ttactgccat tgaaggccat gtgcattagt
     2941 cccaagttac acattaaaaa gtcacatgta gcttaccaac atcagtgctg ttcaagcaca
     3001 gcctcatcta ctattcaaac tgtggcacca tctaaaatat gccagaattt ttttatttaa
     3061 tgaatttgac cctgaaatat gtattaatat cactcctgtg atttttttgt aatcagctta
     3121 caattacagg aatgcaagcc tgattcatta caagtttcac tacactttct ctgacaacat
     3181 cacctactga actcagacca gctagttgct ccttaagtat acaatcatgt cagtaatcct
     3241 catttcaatg aaaaataccc gtattgtact tggtacttgg tagataacca cagagcagta
     3301 ttatgccatt attgtgaata caataagagg taaatgacct acagagctgc tgctgctgtt
     3361 gtgttagatt gtaaacacag cacaggatca aggaggtgtc catcactatg accaatacta
     3421 gcactttgca caggctcttt gaaaggctga aaagagcctt attggcgtta tcacaacaaa
     3481 atacgcaaat acggaaaaca acgtattgaa cttcgcaaac aaaaaacagc gattttgatg
     3541 aaaatcgctt aggccttgct cttcaaacaa tccagcttct ccttctttca ctctcaagtt
     3601 gcaagaagca agtgtagcaa tgtgcacgcg acagccgggt gtgtgacgct ggaccaatca
     3661 gagcgcagag ctccgaaagt ttacctttta tggctagagc cggcatctgc cgtcatataa
     3721 aagagcgcgc ccagcgtctc agcctcactt tgagctcctc cacacgcagc tagtgcggaa
     3781 tatcatctgc ctgtaaccca ttctctaaag tcgacaaacc cccccaaacc taaggtgagt
     3841 tgatctttaa gctttttaca ttttcagctc gcatatatca attcgaacgt ttaattagaa
     3901 tgtttaaata aagctagatt aaatgattag gctcagttac cggtcttttt tttctcattt
     3961 acgtgcgaac tctgcttaaa ctctagttat tctttattaa tatgtggtta tttttatata
     4021 tgtatgttat cataactgta ctggctatgt caggtggtaa tgactgtaac gttacgttac
     4081 tcgttgtagg cacgacattg aatgggccgg tgttgaaata agtcttcaac cccttttaac
     4141 ctcaaaatgt gctctggtta acaaggattt taacagctat cagtatgact gtgcggtttt
     4201 aaagccgtta gtgaggcacg ttgcacactt gatggatggc cggaatggga agttctttat
     4261 gcaggcagtg ctgcagcagg gtgtgaccta ctttagctaa cgttagccgg ctaaccagca
     4321 ttcatctgcc ggtaacttga gtctaatatt ctctatgtga tatcgaagtg atcaaagaca
     4381 cgtctgttag ctcactttaa ccaactgtag tgaaaaatag cgcagtgtgc agcccttcaa
     4441 gtctttcatt taggctgatt attcaatcat tttattaact attaacgcgt tactaaacgt
     4501 aaggtaacgt agtcagtttt taataactgg tgaaaagtac tggttgggtt taaatggtga
     4561 cttataattg tgttggaggg ggaaaccttt ttgataaagg ctatataatc tcaaatgaat
     4621 gggctgagga tggtgttcac aggtgcttta gtgaagtccg ctcgtgaaga gtcgctgaag
     4681 tgactgcaga tctgtagcgc atgcgttttg gcagacggcc gttgaaattc ggttgagtaa
     4741 ttgataccag gtgaggctag aggatgtaga aattcatttg tgtagaattt agggagtggc
     4801 ctggcgtgat gaatgtcgaa atccgttcct ttttactgaa ccctatgtct ctgctgagtg
     4861 ccacaccgcc ggcacaaagc gtctcaaacc attgcctttt atggtaataa tgagaatgca
     4921 gagggacttc ctttgtctgg cacatctgag gcgcgcattg tcacactagc acccactagc
     4981 ggtcagactg cagacaaaca ggaagctgac tccacatggt cacatgctca ctgaagtgtt
     5041 gacttccctg acagctgtgc actttctaaa ccggttttct cattcattta cagttcagcc
     5101 ggaattcacc tgcaggtgga tccatctaga actactagag gatccccggg taccggtaga
     5161 aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattaaatg
     5221 gtgatgttaa tgggcacaaa tttcctgtca gtggagaggg tgaaggtgat gcaacatacg
     5281 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac
     5341 ttgtcactac tctcgcgtat ggtcttcaat gctttgcgag atacccagat catatgaaac
     5401 agcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
     5461 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
     5521 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
     5581 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
     5641 gaatcaaagt taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag
     5701 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     5761 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc
     5821 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataaa
     5881 tgtccagacc tgcaggaatt cgatatcaag cttatcgatg aacaaacccc cccaaaccta
     5941 aggtgagttg atctttaagc tttttacatt ttcagctcgc atatatcaat tcgaacgttt
     6001 aattagaatg tttaaataaa gctagattaa atgattaggc tcagttaccg gtcttttttt
     6061 tctcatttac actagctaag catgctgata acttcgtata gcatacatta tacgaagtta
     6121 ttaccctgtt atccctacaa attaaactgg gcatcagcgc aattcaattg gtttggtaat
     6181 agcaagggaa aatagaatga agtgatctcc aaaaaataag tactttttga ctgtaaataa
     6241 aattgtaagg agtaaaaagt actttttttt ctaaaaaaat gtaattaagt aaaagtaaaa
     6301 gtattgattt ttaattgtac tcaagtaaag taaaaatccc caaaaataat acttaagtac
     6361 agtaatcaag taaaattact caagtacttt acacctctgg gcccaattcg ccctatagtg
     6421 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     6481 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     6541 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     6601 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     6661 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     6721 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     6781 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     6841 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     6901 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     6961 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     7021 ttttaacaaa atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg
     7081 tgcggtattt cacaccgcat caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     7141 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     7201 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     7261 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     7321 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     7381 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     7441 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     7501 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     7561 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     7621 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     7681 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7741 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7801 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7861 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7921 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7981 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     8041 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     8101 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     8161 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     8221 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     8281 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     8341 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     8401 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     8461 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     8521 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     8581 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8641 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8701 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8761 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8821 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8881 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8941 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     9001 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     9061 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagc
//
