LOCUS       62056_11335        5984 bp DNA     circular SYN 25-AUG-2020
DEFINITION  Cloning vector pGBT-RP8.3, complete sequence.
ACCESSION   MT815592
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5984)
  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 5984)
  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 5984)
  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..5984
                     /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             451..1122
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     misc_feature    1134..2309
                     /note="contains 3'UTR, poly(A) signal, and putative
                     boundary element from ocean pout (FRM- Pat Gibbs)"
     regulatory      2353..2713
                     /label=Xenopus gamma-crystallin
                     /note="Xenopus gamma-crystallin"
                     /regulatory_class="promoter"
     misc_feature    2742..2765
                     /note="derived from carp; contains beta actin non-coding
                     exon 1"
     misc_feature    2766..2963
                     /note="derived from carp; contains beta actin intron 1"
     primer_bind     complement(2954..2981)
                     /label=MISC-bact_intron-R1
                     /note="MISC-bact_intron-R1"
     CDS             3014..3712
                     /label=TagBFP
                     /note="monomeric blue fluorescent protein"
     misc_feature    3734..3861
                     /note="derived from carp; contains beta actin splice donor
                     from exon/intron 1 boundary"
     protein_bind    complement(3881..3914)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        3948..3966
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     repeat_region   3971..4188
                     /label=Tol2
                     /note="Tol2"
     CDS             4253..5059
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5205..5793
                     /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 cagccggaat tcacccgggt accggaaggt gcctcctccg aggacgtcat caaggagttc
      481 atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      541 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
      601 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg ctccaaggcc
      661 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
      721 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      781 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
      841 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcggatgtac
      901 cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa ggacggcggc
      961 cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca gctgcccggc
     1021 gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta caccatcgtg
     1081 gaacagtacg agcgcgccga gggccgccac tccaccggag cctaagacta gtaattaagt
     1141 ctcagccacc gttaactgaa catgtcaaaa cctgtggaga ctgttgagat ttgatgttct
     1201 gaaaagataa agcctataaa taaaatgttg cccaaatttc ctgcctgatg tttttctttg
     1261 tctttgctac atggctttgc tgctcggatc ggctcactct gtgtatgcca cgttcacttt
     1321 gtactctcct tctcacggta ggtttattat ttttagatgt gcagttagtt tctgtgaaat
     1381 aacacaccac acactgatat tgtctgtgca ttgacttggt gagtgcacat tgtttttgat
     1441 cttgacatat ttatatttga ttgatcaggt gaactgtgtg aatctaaagt gctccataca
     1501 gatgttctgc attgaaaata ttctcatttt attagtggaa gtgagtgtat gccacatcca
     1561 atcaatttca gcaaacaccc cagtatgatt taatgcaaaa aaatgaaggt atcaaacacg
     1621 cattactact ttgcagttaa atatttaaca tttattccaa cacgaaaaaa agcagtaaat
     1681 aacactttga caaacacgtc aggacatctt atttttgtca ccctcacagg caatttagta
     1741 taatatatta tatatatata tatatcatat aataatattc agtataatat atatatatat
     1801 atcatattat aatattcagt ataatataaa acacaaacac atatatgtat aatataatat
     1861 aacattttta tttattgaga tgcctctatg gaccgtgtta taagaagtaa agatcaggag
     1921 aagtaaacat gaagtgtaat tatgaatact gatgttaaat taagctatga tgagttttca
     1981 ctgttaattt accatctcaa ttaaatgttg atgcctccat gaccaagtta agcagatgag
     2041 actgagacaa ctgtagaaga caagatgttc actttgctga atatagctgg cttgacagtt
     2101 atctatgact ctataaatat atatatattt ttttttttat aaaatgattt atttataact
     2161 atatatccat ttctcagaca ggtgcttcat atccctcact cccgtagctg tccatgctgg
     2221 atctgtcccc gttgttttta aaaagctaaa taagttatta acatgactgc atccagcgag
     2281 ccaaacctgt ctggtgtaca gctaccagag aagcttgaga tcctagtcac cggcgttcct
     2341 agttctttaa actgttacaa agtgtttgga ccagcaactg aattgcactg taaattacca
     2401 catcagggtc tccggggtca acgctgctaa gcaatggact ggaaaatttg tgacattgag
     2461 tttttggaat aactgggaat gctggataac tctttgggta aagctatgga acaaaagtac
     2521 cagcatgaac taagaaatga ctgtcacagc acaacatcaa acacaatgca agacaaaggt
     2581 tctctgcttt tgccttactt gacagaaaaa gtaagctcta ggttcatcgc atgcagacag
     2641 cagcaataca tgacatatat aaaggctggc tcttttctcc cttccattga actgaaactt
     2701 ccactcagtc agacttgcgg gggatccaca gcctgactaa ctcgacaaac ccccccaaac
     2761 ctaaggtgag ttgatcttta agctttttac attttcagct cgcatatatc aattcgcgcg
     2821 cattgtcaca ctagcaccca ctagcggtca gactgcagac aaacaggaag ctgactccac
     2881 atggtcacat gctcactgaa gtgttgactt ccctgacagc tgtgcacttt ctaaaccggt
     2941 tttctcattc atttacagtt cagccggaat tcacctcgag accatggctc caaagaagaa
     3001 gcgtaaggta accatgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     3061 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     3121 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     3181 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     3241 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     3301 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     3361 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     3421 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     3481 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     3541 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     3601 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     3661 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctca attaaagctg
     3721 agatctacct aaggtgagtt gatctttaag ctttttacat tttcagctcg catatatcaa
     3781 ttcgaacgtt taattagaat gtttaaataa agctagatta aatgattagg ctcagttacc
     3841 ggtctttttt ttctcattta cactgagctc aagacgtctg ataacttcgt atagcataca
     3901 ttatacgaag ttattaccct gttatcccta tggctagcga tgggatctaa tacgactcac
     3961 tatagggaga ccaagggaaa atagaatgaa gtgatctcca aaaaataagt actttttgac
     4021 tgtaaataaa attgtaagga gtaaaaagta cttttttttc taaaaaaatg taattaagta
     4081 aaagtaaaag tattgatttt taattgtact caagtaaagt aaaaatcccc aaaaataata
     4141 cttaagtaca gtaatcaagt aaaattactc aagtacttta cacctctggg atccagctgc
     4201 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     4261 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     4321 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     4381 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     4441 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     4501 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     4561 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     4621 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     4681 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     4741 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     4801 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     4861 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     4921 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     4981 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     5041 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcaaaaagg
     5101 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5161 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5221 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5281 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5341 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5401 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     5461 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     5521 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     5581 tacctacagc gtgagctatg agaaagcgcc acggttcccg aagggagaaa ggcggacagg
     5641 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5701 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5761 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5821 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5881 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5941 gagcgcagcg agtcagtgag cgaggaagcg gaacagattt aaat
//
