LOCUS       40924_21200        8392 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-R14, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8392)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     In vivo protein trapping produces a functional expression codex of 
            the vertebrate proteome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8392)
  AUTHORS   Clark KJ, Balciunas D, Pagoda H-M., Ding Y, Westcot SE, Bedell VM, 
            Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielson A, 
            Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-OCT-2010) Biochemistry and Molecular Biology, Mayo 
            Clinic, 221 4th Ave. SW, Rochester, MN 55902, USA
REFERENCE   3  (bases 1 to 8392)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8392)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (01-OCT-2010) Biochemistry and Molecular Biology, Mayo Clinic, 221 
            4th Ave. SW, Rochester, MN 55902, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  complement(2..716)
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="Tol2 long ITR-R"
     protein_bind    734..767
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     intron          811..908
                     /note="carp beta-actin intron 1 splice acceptor"
     CDS             941..1612
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     rep_origin      1588..2044
                     /label=F1 Ori
                     /note="F1 Ori"
     regulatory      1625..2629
                     /label=Danio rerio beta-actin 3'UTR and poly(A) signal
                     /note="Danio rerio beta-actin 3'UTR and poly(A) signal"
                     /regulatory_class="polyA_signal_sequence"
     misc_recomb     2726..2759
                     /label=loxP
                     /note="loxP"
     protein_bind    complement(2726..2759)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     mobile_element  2780..5453
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="Tol2 long ITR-L"
     promoter        complement(5466..5484)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5491..5507)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5648..6103
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6181..6285
                     /label=AmpR promoter
     CDS             6286..7143
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      7317..7905
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8193..8214
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        8229..8259
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8267..8283
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     8291..8307
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8325..8343
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gcagaggtgt aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa
       61 aattttactc aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa
      121 aagtactcca ttaaaattgt acttgagtat taaggaagta aaagtaaaag caagaaagaa
      181 aactagagat tcttgtttaa gcttttaatc tcaaaaaaca ttaaatgaaa tgcatacaag
      241 gttttatcct gctttagaac tgtttgtatt taattatcaa actataagac agacaatcta
      301 atgccagtac acgctactca aagttgtaaa acctcagatt taacttcagt agaagctgat
      361 tctcaaaatt gttagtgtca agcctagctc ttttggggct gaaaagcaat cctgcagtgc
      421 tgaaaagcct ctcacaggca gccgatgcgg gaagaggtgt attagtcttg atagagaggc
      481 tgcaaatagc aggaaacgtg agcagagact ccctggtgtc tgaaacacag gcggccgctt
      541 gagactaggt taagtaagtg acaggcacac ttttgatgtt ttggtagctg gaattcgaag
      601 ccatcgtctt ggtccaggac tcgtgaggca tcttggaatt ccacaccatc acttccctca
      661 ccacagcctt cagaatcagt gtcatcactt tcacaccttc ttgcctcagt ctcgatgtag
      721 ggataacagg gtaataactt cgtataatgt atgctatacg aagttatcag catgccaccc
      781 actagcggtc agactgcaga ttgcagcacg aaacaggaag ctgactccac atggtcacat
      841 gctcactgaa gtgttgactt ccctgacagc tgtgcacttt ctaaaccggt tttctcattc
      901 atttacagtt cagccggaat tcacccgggt accggaaggt gcctcctccg aggacgtcat
      961 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga
     1021 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     1081 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg
     1141 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     1201 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     1261 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     1321 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     1381 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa
     1441 ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca
     1501 gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta
     1561 caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg cctaagaaat
     1621 cgatacggac tgttaccact tcacgccgac tcaaactgcg cagagaaact tcaaacgaca
     1681 acattggcat ggcttttgtt atttttggcg cttgactcag gatctaaaaa ctggaacggt
     1741 gaaggtgacg gcaatgtttt ttggcaaata agcatccccg aagttctaca atgcatctga
     1801 ggactccatg ttttgttttc tttagtcatt ccaaatgttt gttaaatgca ttgttccgaa
     1861 acttatttgc ctctatgaag gctgcccagt aattgggagc atacttaaca ttgtagtatt
     1921 gtatgtaaat tatgtaacaa aacaatgtct gggtttttgt acttttcagc cttaaaatct
     1981 tgggtttatt tttttttctt tgttccaaaa aaacgaagct ttaccattca agatgtaaag
     2041 gtttccattc cccctgggca tattgtaaaa gctgtgtgga acgtggcggt gccagacatt
     2101 tggtggggcc aacctgtaca ctgactaatt caattccaat aaaagtgcac atgtaagaca
     2161 tcctacccct gtctcatttc ctgtcttggc tctgagtggc ttcaataaaa atggcttttg
     2221 tgaagttgca ccatgttaat tacatttcag cactttcaca ccaggttttt gactaatttg
     2281 aatttatcga agtttgacct tttttttttt ttttccttca agctaatttg aggttatcaa
     2341 accatttcat tggaggaaac ttaatgttta aaagcataaa ttcgaaacat gattcattta
     2401 tttaactgct taatggtgaa acaaacttca agtaatacac caagtgttcc ctcatatgaa
     2461 ggtagctaac tgcaaacggc cttaactttc catcttaagt tgagctacat aaaggcttta
     2521 acaggaagga agagccagtt taaaagtcaa accaccatga ctgatataaa ttcttgtcca
     2581 tataaccttg cgtttcacac aatatagcta tctactgcta aataaattat cgcgattcct
     2641 attctctaga aagtatagga acttcctagg agcgctccgg agatcttcct attctctaga
     2701 aagtatagga acttcttaat taacgataac ttcgtataat gtatgctata cgaagttata
     2761 ttaccctgtt atccctacca tgatcattca tggcactggc cagtacctca aaagtatgag
     2821 agcccattaa tcttttgcag gcaagtgcag cggaatgtct ttcaagactt ccagggttga
     2881 tccagtgagc agttacacca atgaatgact ttctacgtgc agtccaacaa tccgttgtgg
     2941 ttgcaatcca ttcaacttca ctcatggcag cagtcacttt ctgtttcatg atcagagcag
     3001 cttcagctat cttggagcgt aaagtaggcc ttgtaatgac agaaatgcca ggctgcagtg
     3061 tactaatcag ctctttaaat gatggcagat caacagtgct gaaaggatga agtccttgaa
     3121 tgatgtacct taatatagct ttgttcacag tgactggaga cacatgtttg actgggaaaa
     3181 ctgagtcaac tttcagttgc ttactgctgg aagcatgggt ggaggtcccg atctttctct
     3241 tctgtgctgt caatttagag tagtttttga ggtaatttgg gtgcattctc tgtaaaaaca
     3301 acaaataaat atttatattg tatgcaataa atgtaacaat tacacataca ctctctgatt
     3361 acagcattaa agggttagtt cacccaaaaa tgaaaagatg tcattaatga ctcaccctca
     3421 tgtcgttcca agcccgtaac acctccgttc atcttcggaa cacagtttaa gatattttag
     3481 atttagtccg agagctttct gtccctccac tgaaaatgta tgtacggtat actgtccatg
     3541 tccagaaagg taataaaaac atcaaagtag tccatgtgac atcagtgggt taggtagaat
     3601 tatttgaagc atcgaaaata cattttggtc caaaaataac aaaacctacg actttattca
     3661 gcattgtagc gtcactgcga agtcgtgaac gcggattgac aacagacccg gaagagaata
     3721 caatgccgaa taaagtcgta ggttttgtta tttttggacc aaaatgtatt cgatgcttca
     3781 aaaaattcta actaacccac tgatgtcaca tggactactt tgatgttttt attacctttc
     3841 tggacatgga cagtataccg tacatacatt ctcaatggag gcacagaaag ctctcggact
     3901 aaatctaaaa tatcttaaac tgtgttctga agatgaacgg aggtcttacg ggcttggaac
     3961 gacatgaggg cgagtcatta atgacattat tttcattttt gggtgaacta accctttaat
     4021 aaaactgaca tttgcagcca attgcacact ttataatgtt aaatacttgt tgattttaga
     4081 aatacagtag catttagtga caatcaaaca ttaattttat ttctgattat attgacaaaa
     4141 acacaggact gtaaggctga catgacaggg tgaattgatt tcattacatt agttttgatg
     4201 gtaacacttt tacaataagg ttcgttagtt aactacatta gttaacatga actaataatg
     4261 aactgcacgt atacatgtta atttcaacat ttactaatac tttatatctt gttaacatta
     4321 gttaatgcac tgtgaactaa catgaacaaa caatgaacat ctgtatttct attaactaac
     4381 gttaacaaag attaacagat acagtaacaa atgtattgct catggttatt taatgttagt
     4441 taatacatga actaatcaac ctcattgtaa agtgttaccg ttttgacaat acaacatgtt
     4501 tggaaaaaaa tttctcgctt ggttttcttg tgttttgcat gaattcaaac acaatgagag
     4561 cacggaatga taatgattat tcaaagctgc aataggcgcc aaatttcccg tggacctaac
     4621 gttaccaatt acaaatgcat tttagcattt tattacgtga gccttatatt gtcaatcgca
     4681 tgcattagtg aaaataggcc tactacacat ataaaaggcg cgccatcaac gtcaaaacat
     4741 gcacacccaa aaaaaaaaaa aaaaaaaaaa aaaaaaacta tcagtaaaac aaaatactta
     4801 atgtacttac ctcaatatgc ttccttaggt ttgatggcga acttttgaag gccgatattt
     4861 ctttattaag cgggagacag aggacacatt tcatcttgaa tgaatcttta tttacaccac
     4921 ttaaagaaaa gaattcgcga agatacggcc acgggtgctc ttgatcctgt ggctgatttt
     4981 ggactgtgct gctcgcagct gctgatgaat cacatacttc ctccattttc ttccactgat
     5041 tgactgttat aatttcccta atttccaggt caaggtgctg tgcattgtgg taatagatgt
     5101 gacatgacgt cacttccaaa ggaccaatga acatgtctga ccaatttcat ataatgtgaa
     5161 aacgattttc ataggcagaa taaataacat ttaaattaaa ctgggcatca gcgcaattca
     5221 attggtttgg taatagcaag ggaaaataga atgaagtgat ctccaaaaaa taagtacttt
     5281 ttgactgtaa ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt
     5341 aagtaaaagt aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa
     5401 taatacttaa gtacagtaat caagtaaaat tactcaagta ctttacacct ctgggcccaa
     5461 ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg
     5521 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5581 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5641 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5701 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5761 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5821 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5881 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5941 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     6001 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     6061 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     6121 tccttacgca tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg
     6181 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6241 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6301 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6361 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6421 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6481 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6541 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6601 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6661 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6721 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6781 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6841 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6901 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6961 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7021 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7081 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7141 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7201 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7261 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7321 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7381 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7441 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7501 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7561 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7621 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7681 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7741 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7801 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7861 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7921 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     7981 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8041 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     8101 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8161 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8221 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8281 caatttcaca caggaaacag ctatgaccat gattacgcca agctatttag gtgacactat
     8341 agaatactca agctatgcat ccaacgcgtt gggagctctc ccatatggtc ga
//
