LOCUS       Exported                9118 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 9118)
  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..9118
                     /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             652..1326
                     /label=mRFP*
                     /note="mRFP with AUG removed"
     polyA_signal    1339..2514
                     /label=Ocean Pout Terminator
                     /note="Ocean pout antifreeze protein 3'UTR, poly(A) signal,
                     and putative boundary element (Pat Gibbs)"
     misc_feature    2603..3738
                     /label=beta-actin promoter
                     /note="Carp beta-actin promoter"
     exon            3744..3835
                     /label=beta actin exon 1
                     /note="carp beta actin exon 1 (non-coding)"
     intron          3836..5099
                     /label=beta-actin intron 1
                     /note="carp beta actin intron 1"
     CDS             5164..5880
                     /label=GFP
                     /note="GM2 variant"
     misc_feature    5944..6071
                     /label=SD
                     /note="carp beta actin splice-donor exon/intron 1 boundary"
     misc_recomb     6089..6122
                     /label=loxP
     misc_feature    6121..6138
                     /label=I-SceI
     misc_feature    6139..6400
                     /label=Tol2_ITR-L
     rep_origin      6595..7050
                     /label=F1 Ori
     CDS             complement(7233..8093)
                     /label=AmpR
     rep_origin      complement(8154..9034)
                     /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 tcagccggaa ttcacccggg taccggaagg tgcctcctcc
      661 gaggacgtca tcaaggagtt catgcgcttc aaggtgcgca tggagggctc cgtgaacggc
      721 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
      781 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtcccctcag
      841 ttccagtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
      901 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
      961 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     1021 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     1081 gcctccaccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagatgagg
     1141 ctgaagctga aggacggcgg ccactacgac gccgaggtca agaccaccta catggccaag
     1201 aagcccgtgc agctgcccgg cgcctacaag accgacatca agctggacat cacctcccac
     1261 aacgaggact acaccatcgt ggaacagtac gagcgcgccg agggccgcca ctccaccggc
     1321 gcctaagaaa tcctagtaat taagtctcag ccaccgttaa ctgaacatgt caaaacctgt
     1381 ggagactgtt gagatttgat gttctgaaaa gataaagcct ataaataaaa tgttgcccaa
     1441 atttcctgcc tgatgttttt ctttgtcttt gctacatggc tttgctgctc ggatcggctc
     1501 actctgtgta tgccacgttc actttgtact ctccttctca cggtaggttt attattttta
     1561 gatgtgcagt tagtttctgt gaaataacac accacacact gatattgtct gtgcattgac
     1621 ttggtgagtg cacattgttt ttgatcttga catatttata tttgattgat caggtgaact
     1681 gtgtgaatct aaagtgctcc atacagatgt tctgcattga aaatattctc attttattag
     1741 tggaagtgag tgtatgccac atccaatcaa tttcagcaaa caccccagta tgatttaatg
     1801 caaaaaaatg aaggtatcaa acacgcatta ctactttgca gttaaatatt taacatttat
     1861 tccaacacga aaaaaagcag taaataacac tttgacaaac acgtcaggac atcttatttt
     1921 tgtcaccctc acaggcaatt tagtataata tattatatat atatatatat catataataa
     1981 tattcagtat aatatatata tatatatcat attataatat tcagtataat ataaaacaca
     2041 aacacatata tgtataatat aatataacat ttttatttat tgagatgcct ctatggaccg
     2101 tgttataaga agtaaagatc aggagaagta aacatgaagt gtaattatga atactgatgt
     2161 taaattaagc tatgatgagt tttcactgtt aatttaccat ctcaattaaa tgttgatgcc
     2221 tccatgacca agttaagcag atgagactga gacaactgta gaagacaaga tgttcacttt
     2281 gctgaatata gctggcttga cagttatcta tgactctata aatatatata tatttttttt
     2341 tttataaaat gatttattta taactatata tccatttctc agacaggtgc ttcatatccc
     2401 tcactcccgt agctgtccat gctggatctg tccccgttgt ttttaaaaag ctaaataagt
     2461 tattaacatg actgcatcca gcgagccaaa cctgtctggt gtacagctac cagagaagct
     2521 tgagatccta ggaagttcct atactttcta gagaatagga atcgcgagtg ctagcaatta
     2581 accctcacta aagggaacaa aagcttttag accttcttac ttttggggat tatataagta
     2641 ttttctcaat aaatatctca tatcttactg tggtttaact gctgaatcta aaattttaat
     2701 acaaaagtag ttatatttgt tgtacattgt aaactataac ttaacttcag tttcagagaa
     2761 actcatgtgc tcaaaatgta aaaaaagttt cctgttaaat attttgtaaa tgtattgaag
     2821 acaaaataag aaaaaaaaaa atataagcca ctaaatcaca ctgtccttgg tatcagcaag
     2881 agattctgac ataatcagct gtttttgttt attactgcca ttgaaggcca tgtgcattag
     2941 tcccaagtta cacattaaaa agtcacatgt agcttaccaa catcagtgct gttcaagcac
     3001 agcctcatct actattcaaa ctgtggcacc atctaaaata tgccagaatt tttttattta
     3061 atgaatttga ccctgaaata tgtattaata tcactcctgt gatttttttg taatcagctt
     3121 acaattacag gaatgcaagc ctgattcatt acaagtttca ctacactttc tctgacaaca
     3181 tcacctactg aactcagacc agctagttgc tccttaagta tacaatcatg tcagtaatcc
     3241 tcatttcaat gaaaaatacc cgtattgtac ttggtacttg gtagataacc acagagcagt
     3301 attatgccat tattgtgaat acaataagag gtaaatgacc tacagagctg ctgctgctgt
     3361 tgtgttagat tgtaaacaca gcacaggatc aaggaggtgt ccatcactat gaccaatact
     3421 agcactttgc acaggctctt tgaaaggctg aaaagagcct tattggcgtt atcacaacaa
     3481 aatacgcaaa tacggaaaac aacgtattga acttcgcaaa caaaaaacag cgattttgat
     3541 gaaaatcgct taggccttgc tcttcaaaca atccagcttc tccttctttc actctcaagt
     3601 tgcaagaagc aagtgtagca atgtgcacgc gacagccggg tgtgtgacgc tggaccaatc
     3661 agagcgcaga gctccgaaag tttacctttt atggctagag ccggcatctg ccgtcatata
     3721 aaagagcgcg cccagcgtct cagcctcact ttgagctcct ccacacgcag ctagtgcgga
     3781 atatcatctg cctgtaaccc attctctaaa gtcgacaaac ccccccaaac ctaaggtgag
     3841 ttgatcttta agctttttac attttcagct cgcatatatc aattcgaacg tttaattaga
     3901 atgtttaaat aaagctagat taaatgatta ggctcagtta ccggtctttt ttttctcatt
     3961 tacgtgcgaa ctctgcttaa actctagtta ttctttatta atatgtggtt atttttatat
     4021 atgtatgtta tcataactgt actggctatg tcaggtggta atgactgtaa cgttacgtta
     4081 ctcgttgtag gcacgacatt gaatgggccg gtgttgaaat aagtcttcaa ccccttttaa
     4141 cctcaaaatg tgctctggtt aacaaggatt ttaacagcta tcagtatgac tgtgcggttt
     4201 taaagccgtt agtgaggcac gttgcacact tgatggatgg ccggaatggg aagttcttta
     4261 tgcaggcagt gctgcagcag ggtgtgacct actttagcta acgttagccg gctaaccagc
     4321 attcatctgc cggtaacttg agtctaatat tctctatgtg atatcgaagt gatcaaagac
     4381 acgtctgtta gctcacttta accaactgta gtgaaaaata gcgcagtgtg cagcccttca
     4441 agtctttcat ttaggctgat tattcaatca ttttattaac tattaacgcg ttactaaacg
     4501 taaggtaacg tagtcagttt ttaataactg gtgaaaagta ctggttgggt ttaaatggtg
     4561 acttataatt gtgttggagg gggaaacctt tttgataaag gctatataat ctcaaatgaa
     4621 tgggctgagg atggtgttca caggtgcttt agtgaagtcc gctcgtgaag agtcgctgaa
     4681 gtgactgcag atctgtagcg catgcgtttt ggcagacggc cgttgaaatt cggttgagta
     4741 attgatacca ggtgaggcta gaggatgtag aaattcattt gtgtagaatt tagggagtgg
     4801 cctggcgtga tgaatgtcga aatccgttcc tttttactga accctatgtc tctgctgagt
     4861 gccacaccgc cggcacaaag cgtctcaaac cattgccttt tatggtaata atgagaatgc
     4921 agagggactt cctttgtctg gcacatctga ggcgcgcatt gtcacactag cacccactag
     4981 cggtcagact gcagacaaac aggaagctga ctccacatgg tcacatgctc actgaagtgt
     5041 tgacttccct gacagctgtg cactttctaa accggttttc tcattcattt acagttcagc
     5101 cggaattcac ctgcaggtgg atccatctag aactactaga ggatccccgg gtaccggtag
     5161 aaaatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattaaat
     5221 ggtgatgtta atgggcacaa atttcctgtc agtggagagg gtgaaggtga tgcaacatac
     5281 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca
     5341 cttgtcacta ctctcgcgta tggtcttcaa tgctttgcga gatacccaga tcatatgaaa
     5401 cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt
     5461 ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg tgataccctt
     5521 gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat tcttggacac
     5581 aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa acaaaagaat
     5641 ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca
     5701 gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat
     5761 tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc
     5821 cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaataa
     5881 atgtccagac ctgcaggaat tcgatatcaa gcttatcgat gaacaaaccc ccccaaacct
     5941 aaggtgagtt gatctttaag ctttttacat tttcagctcg catatatcaa ttcgaacgtt
     6001 taattagaat gtttaaataa agctagatta aatgattagg ctcagttacc ggtctttttt
     6061 ttctcattta cactagctaa gcatgctgat aacttcgtat agcatacatt atacgaagtt
     6121 attaccctgt tatccctaca aattaaactg ggcatcagcg caattcaatt ggtttggtaa
     6181 tagcaaggga aaatagaatg aagtgatctc caaaaaataa gtactttttg actgtaaata
     6241 aaattgtaag gagtaaaaag tacttttttt tctaaaaaaa tgtaattaag taaaagtaaa
     6301 agtattgatt tttaattgta ctcaagtaaa gtaaaaatcc ccaaaaataa tacttaagta
     6361 cagtaatcaa gtaaaattac tcaagtactt tacacctctg ggcccaattc gccctatagt
     6421 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     6481 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     6541 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     6601 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     6661 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     6721 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     6781 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     6841 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     6901 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6961 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     7021 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     7081 gtgcggtatt tcacaccgca tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     7141 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     7201 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     7261 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     7321 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     7381 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     7441 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     7501 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     7561 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     7621 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     7681 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     7741 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     7801 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     7861 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     7921 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     7981 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     8041 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     8101 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     8161 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     8221 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     8281 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     8341 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     8401 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     8461 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     8521 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     8581 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     8641 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     8701 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     8761 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     8821 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     8881 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     8941 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     9001 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     9061 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagc
//
