LOCUS       62056_11330        5985 bp DNA     circular SYN 25-AUG-2020
DEFINITION  Cloning vector pGBT-RP8.2, complete sequence.
ACCESSION   MT815591
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5985)
  AUTHORS   Ichino N, Serres MR, Urban RM, Urban MD, Treichel AJ, Schaefbauer 
            KJ, Greif LE, Varshney GK, Skuster KJ, McNulty MS, Daby CL, Wang Y, 
            Liao HK, El-Rass S, Ding Y, Liu W, Anderson JL, Wishman MD, 
            Sabharwal A, Schimmenti LA, Sivasubbu S, Balciunas D, Hammerschmidt 
            M, Farber SA, Wen XY, Xu X, McGrail M, Essner JJ, Burgess SM, Clark 
            KJ, Ekker SC.
  TITLE     Building the vertebrate codex using the gene breaking protein trap 
            library
  JOURNAL   Elife 9 (2020) In press
  PUBMED    32779569
REFERENCE   2  (bases 1 to 5985)
  AUTHORS   Ichino N, Serres MR, Urban RM, Urban MD, Treichel AJ, Schaefbauer 
            KJ, Greif LE, Varshney GK, Skuster KJ, McNulty MS, Daby CL, Wang Y, 
            Liao HK, El-Rass S, Ding Y, Liu W, Anderson JL, Wishman MD, 
            Sabharwal A, Schimmenti LA, Sivasubbu S, Balciunas D, Hammerschmidt 
            M, Farber SA, Wen XY, Xu X, McGrail M, Essner JJ, Burgess SM, Clark 
            KJ, Ekker SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUL-2020) Biochemistry and Molecular Biology, Mayo 
            Clinic, 200 First Street SW, Rochester, Olmsted, MN 55905, USA
REFERENCE   3  (bases 1 to 5985)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife 9 
            (2020) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-JUL-2020) Biochemistry and Molecular Biology, Mayo Clinic, 200 
            First Street SW, Rochester, Olmsted, MN 55905, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..196
                     /label=Tol2
                     /note="Tol2"
     promoter        complement(204..222)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_recomb     248..281
                     /label=loxP
                     /note="loxP"
     misc_feature    288..397
                     /note="derived from carp; contains beta actin splice
                     acceptor from intron 1/exon2 boundary"
     CDS             452..1123
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     misc_feature    1135..2310
                     /note="contains 3'UTR, poly(A) signal, and putative
                     boundary element from ocean pout (FRM- Pat Gibbs)"
     regulatory      2354..2714
                     /label=Xenopus gamma-crystallin
                     /note="Xenopus gamma-crystallin"
                     /regulatory_class="promoter"
     misc_feature    2743..2766
                     /note="derived from carp; contains beta actin non-coding
                     exon 1"
     misc_feature    2767..2964
                     /note="derived from carp; contains beta actin intron 1"
     primer_bind     complement(2955..2982)
                     /label=MISC-bact_intron-R1
                     /note="MISC-bact_intron-R1"
     CDS             3015..3713
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
     misc_feature    3735..3862
                     /note="derived from carp; contains beta actin splice donor
                     from exon/intron 1 boundary"
     protein_bind    complement(3882..3915)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3949..3967
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   3972..4189
                     /label=Tol2
                     /note="Tol2"
     CDS             4254..5060
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5206..5794
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtaccgagc agaggtgtaa aaagtactca aaaattttac tcaagtgaaa gtacaagtac
       61 ttagggaaaa ttttactcaa ttaaaagtaa aagtatctgg ctagaatctt acttgagtaa
      121 aagtaaaaaa gtactccatt aaaattgtac ttgagtatta aggaagtaaa agtaaaagca
      181 agaaagaaaa ctagagtggt ctccctttag tgagggttaa ttgatgggat agggataaca
      241 gggtaatata acttcgtata gcatacatta tacgaagtta tcgttaccac ccactagcgg
      301 tcagactgca gattgcagca cgaaacagga agctgactcc acatggtcac atgctcactg
      361 aagtgttgac ttccctgaca gctgtgcact ttctaaaccg gttttctcat tcatttacag
      421 tcagccggaa ttcacccggg taccggaagg tgcctcctcc gaggacgtca tcaaggagtt
      481 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
      541 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
      601 cggccccctg cccttcgcct gggacatcct gtcccctcag ttccagtacg gctccaaggc
      661 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
      721 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
      781 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
      841 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccaccg agcggatgta
      901 ccccgaggac ggcgccctga agggcgagat caagatgagg ctgaagctga aggacggcgg
      961 ccactacgac gccgaggtca agaccaccta catggccaag aagcccgtgc agctgcccgg
     1021 cgcctacaag accgacatca agctggacat cacctcccac aacgaggact acaccatcgt
     1081 ggaacagtac gagcgcgccg agggccgcca ctccaccgga gcctaagact agtaattaag
     1141 tctcagccac cgttaactga acatgtcaaa acctgtggag actgttgaga tttgatgttc
     1201 tgaaaagata aagcctataa ataaaatgtt gcccaaattt cctgcctgat gtttttcttt
     1261 gtctttgcta catggctttg ctgctcggat cggctcactc tgtgtatgcc acgttcactt
     1321 tgtactctcc ttctcacggt aggtttatta tttttagatg tgcagttagt ttctgtgaaa
     1381 taacacacca cacactgata ttgtctgtgc attgacttgg tgagtgcaca ttgtttttga
     1441 tcttgacata tttatatttg attgatcagg tgaactgtgt gaatctaaag tgctccatac
     1501 agatgttctg cattgaaaat attctcattt tattagtgga agtgagtgta tgccacatcc
     1561 aatcaatttc agcaaacacc ccagtatgat ttaatgcaaa aaaatgaagg tatcaaacac
     1621 gcattactac tttgcagtta aatatttaac atttattcca acacgaaaaa aagcagtaaa
     1681 taacactttg acaaacacgt caggacatct tatttttgtc accctcacag gcaatttagt
     1741 ataatatatt atatatatat atatatcata taataatatt cagtataata tatatatata
     1801 tatcatatta taatattcag tataatataa aacacaaaca catatatgta taatataata
     1861 taacattttt atttattgag atgcctctat ggaccgtgtt ataagaagta aagatcagga
     1921 gaagtaaaca tgaagtgtaa ttatgaatac tgatgttaaa ttaagctatg atgagttttc
     1981 actgttaatt taccatctca attaaatgtt gatgcctcca tgaccaagtt aagcagatga
     2041 gactgagaca actgtagaag acaagatgtt cactttgctg aatatagctg gcttgacagt
     2101 tatctatgac tctataaata tatatatatt ttttttttta taaaatgatt tatttataac
     2161 tatatatcca tttctcagac aggtgcttca tatccctcac tcccgtagct gtccatgctg
     2221 gatctgtccc cgttgttttt aaaaagctaa ataagttatt aacatgactg catccagcga
     2281 gccaaacctg tctggtgtac agctaccaga gaagcttgag atcctagtca ccggcgttcc
     2341 tagttcttta aactgttaca aagtgtttgg accagcaact gaattgcact gtaaattacc
     2401 acatcagggt ctccggggtc aacgctgcta agcaatggac tggaaaattt gtgacattga
     2461 gtttttggaa taactgggaa tgctggataa ctctttgggt aaagctatgg aacaaaagta
     2521 ccagcatgaa ctaagaaatg actgtcacag cacaacatca aacacaatgc aagacaaagg
     2581 ttctctgctt ttgccttact tgacagaaaa agtaagctct aggttcatcg catgcagaca
     2641 gcagcaatac atgacatata taaaggctgg ctcttttctc ccttccattg aactgaaact
     2701 tccactcagt cagacttgcg ggggatccac agcctgacta actcgacaaa cccccccaaa
     2761 cctaaggtga gttgatcttt aagcttttta cattttcagc tcgcatatat caattcgcgc
     2821 gcattgtcac actagcaccc actagcggtc agactgcaga caaacaggaa gctgactcca
     2881 catggtcaca tgctcactga agtgttgact tccctgacag ctgtgcactt tctaaaccgg
     2941 ttttctcatt catttacagt tcagccggaa ttcacctcga gaccatggct ccaaagaaga
     3001 agcgtaaggt aaccatgagc gagctgatta aggagaacat gcacatgaag ctgtacatgg
     3061 agggcaccgt ggacaaccat cacttcaagt gcacatccga gggcgaaggc aagccctacg
     3121 agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg
     3181 acatcctggc tactagcttc ctctacggca gcaagacctt catcaaccac acccagggca
     3241 tccccgactt cttcaagcag tccttccctg agggcttcac atgggagaga gtcaccacat
     3301 acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca
     3361 tctacaacgt caagatcaga ggggtgaact tcacatccaa cggccctgtg atgcagaaga
     3421 aaacactcgg ctgggaggcc ttcaccgaga cgctgtaccc cgctgacggc ggcctggaag
     3481 gcagaaacga catggccctg aagctcgtgg gcgggagcca tctgatcgca aacatcaaga
     3541 ccacatatag atccaagaaa cccgctaaga acctcaagat gcctggcgtc tactatgtgg
     3601 actacagact ggaaagaatc aaggaggcca acaacgagac ctacgtcgag cagcacgagg
     3661 tggcagtggc cagatactgc gacctcccta gcaaactggg gcacaagctc aattaaagct
     3721 gagatctacc taaggtgagt tgatctttaa gctttttaca ttttcagctc gcatatatca
     3781 attcgaacgt ttaattagaa tgtttaaata aagctagatt aaatgattag gctcagttac
     3841 cggtcttttt tttctcattt acactgagct caagacgtct gataacttcg tatagcatac
     3901 attatacgaa gttattaccc tgttatccct atggctagcg atgggatcta atacgactca
     3961 ctatagggag accaagggaa aatagaatga agtgatctcc aaaaaataag tactttttga
     4021 ctgtaaataa aattgtaagg agtaaaaagt actttttttt ctaaaaaaat gtaattaagt
     4081 aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag taaaaatccc caaaaataat
     4141 acttaagtac agtaatcaag taaaattact caagtacttt acacctctgg gatccagctg
     4201 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
     4261 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
     4321 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
     4381 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     4441 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     4501 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
     4561 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     4621 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     4681 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     4741 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     4801 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     4861 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     4921 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     4981 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     5041 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcaaaaag
     5101 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5161 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5221 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5281 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5341 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5401 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5461 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5521 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5581 atacctacag cgtgagctat gagaaagcgc cacggttccc gaagggagaa aggcggacag
     5641 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5701 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5761 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5821 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     5881 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     5941 cgagcgcagc gagtcagtga gcgaggaagc ggaacagatt taaat
//
