LOCUS       40924_21215        9119 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGBT-RP2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9119)
  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  JOURNAL   Unpublished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 9119)
  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 9119)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9119)
  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            "Unpublished"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..9119
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..43
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        58..88
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    96..112
                     /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     120..136
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        154..172
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     mobile_element  complement(222..413)
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini Tol2 ITR-R"
     misc_recomb     448..481
                     /label=loxP
                     /note="loxP"
     protein_bind    448..481
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          488..620
                     /note="carp beta-actin intron 1 splice acceptor"
     CDS             653..1324
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     regulatory      1340..2515
                     /note="ocean pout antifreeze poly(A) signal; terminator; 
                     putative border element"
                     /regulatory_class="polyA_signal_sequence"
     rep_origin      1588..2044
                     /label=F1 Ori
                     /note="F1 Ori"
     promoter        2577..2595
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2604..3739
                     /label=carp beta-actin promoter region
                     /note="carp beta-actin promoter region"
                     /regulatory_class="promoter"
     exon            3745..3836
                     /note="carp beta-actin exon 1 (non-coding)"
     intron          3837..5100
                     /note="carp beta-actin intron 1"
     CDS             5165..5878
                     /label=GFP
                     /note="green fluorescent protein"
     intron          5945..6072
                     /note="carp beta-actin intron 1 splice donor"
     protein_bind    complement(6090..6123)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  6140..6401
                     /mobile_element_type="transposon:Tol2"
                     /label=ol2
                     /note="mini Tol2 ITR-L"
     promoter        complement(6414..6432)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6439..6455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      6596..7051
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7129..7233
                     /label=AmpR promoter
     CDS             7234..8091
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8265..8853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 ttcagccgga attcacccgg gtaccggaag gtgcctcctc
      661 cgaggacgtc atcaaggagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
      721 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
      781 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccctca
      841 gttccagtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
      901 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
      961 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
     1021 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
     1081 ggcctccacc gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagatgag
     1141 gctgaagctg aaggacggcg gccactacga cgccgaggtc aagaccacct acatggccaa
     1201 gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc aagctggaca tcacctccca
     1261 caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc actccaccgg
     1321 cgcctaagaa atcctagtaa ttaagtctca gccaccgtta actgaacatg tcaaaacctg
     1381 tggagactgt tgagatttga tgttctgaaa agataaagcc tataaataaa atgttgccca
     1441 aatttcctgc ctgatgtttt tctttgtctt tgctacatgg ctttgctgct cggatcggct
     1501 cactctgtgt atgccacgtt cactttgtac tctccttctc acggtaggtt tattattttt
     1561 agatgtgcag ttagtttctg tgaaataaca caccacacac tgatattgtc tgtgcattga
     1621 cttggtgagt gcacattgtt tttgatcttg acatatttat atttgattga tcaggtgaac
     1681 tgtgtgaatc taaagtgctc catacagatg ttctgcattg aaaatattct cattttatta
     1741 gtggaagtga gtgtatgcca catccaatca atttcagcaa acaccccagt atgatttaat
     1801 gcaaaaaaat gaaggtatca aacacgcatt actactttgc agttaaatat ttaacattta
     1861 ttccaacacg aaaaaaagca gtaaataaca ctttgacaaa cacgtcagga catcttattt
     1921 ttgtcaccct cacaggcaat ttagtataat atattatata tatatatata tcatataata
     1981 atattcagta taatatatat atatatatca tattataata ttcagtataa tataaaacac
     2041 aaacacatat atgtataata taatataaca tttttattta ttgagatgcc tctatggacc
     2101 gtgttataag aagtaaagat caggagaagt aaacatgaag tgtaattatg aatactgatg
     2161 ttaaattaag ctatgatgag ttttcactgt taatttacca tctcaattaa atgttgatgc
     2221 ctccatgacc aagttaagca gatgagactg agacaactgt agaagacaag atgttcactt
     2281 tgctgaatat agctggcttg acagttatct atgactctat aaatatatat atattttttt
     2341 ttttataaaa tgatttattt ataactatat atccatttct cagacaggtg cttcatatcc
     2401 ctcactcccg tagctgtcca tgctggatct gtccccgttg tttttaaaaa gctaaataag
     2461 ttattaacat gactgcatcc agcgagccaa acctgtctgg tgtacagcta ccagagaagc
     2521 ttgagatcct aggaagttcc tatactttct agagaatagg aatcgcgagt gctagcaatt
     2581 aaccctcact aaagggaaca aaagctttta gaccttctta cttttgggga ttatataagt
     2641 attttctcaa taaatatctc atatcttact gtggtttaac tgctgaatct aaaattttaa
     2701 tacaaaagta gttatatttg ttgtacattg taaactataa cttaacttca gtttcagaga
     2761 aactcatgtg ctcaaaatgt aaaaaaagtt tcctgttaaa tattttgtaa atgtattgaa
     2821 gacaaaataa gaaaaaaaaa aatataagcc actaaatcac actgtccttg gtatcagcaa
     2881 gagattctga cataatcagc tgtttttgtt tattactgcc attgaaggcc atgtgcatta
     2941 gtcccaagtt acacattaaa aagtcacatg tagcttacca acatcagtgc tgttcaagca
     3001 cagcctcatc tactattcaa actgtggcac catctaaaat atgccagaat ttttttattt
     3061 aatgaatttg accctgaaat atgtattaat atcactcctg tgattttttt gtaatcagct
     3121 tacaattaca ggaatgcaag cctgattcat tacaagtttc actacacttt ctctgacaac
     3181 atcacctact gaactcagac cagctagttg ctccttaagt atacaatcat gtcagtaatc
     3241 ctcatttcaa tgaaaaatac ccgtattgta cttggtactt ggtagataac cacagagcag
     3301 tattatgcca ttattgtgaa tacaataaga ggtaaatgac ctacagagct gctgctgctg
     3361 ttgtgttaga ttgtaaacac agcacaggat caaggaggtg tccatcacta tgaccaatac
     3421 tagcactttg cacaggctct ttgaaaggct gaaaagagcc ttattggcgt tatcacaaca
     3481 aaatacgcaa atacggaaaa caacgtattg aacttcgcaa acaaaaaaca gcgattttga
     3541 tgaaaatcgc ttaggccttg ctcttcaaac aatccagctt ctccttcttt cactctcaag
     3601 ttgcaagaag caagtgtagc aatgtgcacg cgacagccgg gtgtgtgacg ctggaccaat
     3661 cagagcgcag agctccgaaa gtttaccttt tatggctaga gccggcatct gccgtcatat
     3721 aaaagagcgc gcccagcgtc tcagcctcac tttgagctcc tccacacgca gctagtgcgg
     3781 aatatcatct gcctgtaacc cattctctaa agtcgacaaa cccccccaaa cctaaggtga
     3841 gttgatcttt aagcttttta cattttcagc tcgcatatat caattcgaac gtttaattag
     3901 aatgtttaaa taaagctaga ttaaatgatt aggctcagtt accggtcttt tttttctcat
     3961 ttacgtgcga actctgctta aactctagtt attctttatt aatatgtggt tatttttata
     4021 tatgtatgtt atcataactg tactggctat gtcaggtggt aatgactgta acgttacgtt
     4081 actcgttgta ggcacgacat tgaatgggcc ggtgttgaaa taagtcttca acccctttta
     4141 acctcaaaat gtgctctggt taacaaggat tttaacagct atcagtatga ctgtgcggtt
     4201 ttaaagccgt tagtgaggca cgttgcacac ttgatggatg gccggaatgg gaagttcttt
     4261 atgcaggcag tgctgcagca gggtgtgacc tactttagct aacgttagcc ggctaaccag
     4321 cattcatctg ccggtaactt gagtctaata ttctctatgt gatatcgaag tgatcaaaga
     4381 cacgtctgtt agctcacttt aaccaactgt agtgaaaaat agcgcagtgt gcagcccttc
     4441 aagtctttca tttaggctga ttattcaatc attttattaa ctattaacgc gttactaaac
     4501 gtaaggtaac gtagtcagtt tttaataact ggtgaaaagt actggttggg tttaaatggt
     4561 gacttataat tgtgttggag ggggaaacct ttttgataaa ggctatataa tctcaaatga
     4621 atgggctgag gatggtgttc acaggtgctt tagtgaagtc cgctcgtgaa gagtcgctga
     4681 agtgactgca gatctgtagc gcatgcgttt tggcagacgg ccgttgaaat tcggttgagt
     4741 aattgatacc aggtgaggct agaggatgta gaaattcatt tgtgtagaat ttagggagtg
     4801 gcctggcgtg atgaatgtcg aaatccgttc ctttttactg aaccctatgt ctctgctgag
     4861 tgccacaccg ccggcacaaa gcgtctcaaa ccattgcctt ttatggtaat aatgagaatg
     4921 cagagggact tcctttgtct ggcacatctg aggcgcgcat tgtcacacta gcacccacta
     4981 gcggtcagac tgcagacaaa caggaagctg actccacatg gtcacatgct cactgaagtg
     5041 ttgacttccc tgacagctgt gcactttcta aaccggtttt ctcattcatt tacagttcag
     5101 ccggaattca cctgcaggtg gatccatcta gaactactag aggatccccg ggtaccggta
     5161 gaaaatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaaa
     5221 tggtgatgtt aatgggcaca aatttcctgt cagtggagag ggtgaaggtg atgcaacata
     5281 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac
     5341 acttgtcact actctcgcgt atggtcttca atgctttgcg agatacccag atcatatgaa
     5401 acagcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt
     5461 tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct
     5521 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
     5581 caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa
     5641 tggaatcaaa gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc
     5701 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     5761 ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt
     5821 ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata
     5881 aatgtccaga cctgcaggaa ttcgatatca agcttatcga tgaacaaacc cccccaaacc
     5941 taaggtgagt tgatctttaa gctttttaca ttttcagctc gcatatatca attcgaacgt
     6001 ttaattagaa tgtttaaata aagctagatt aaatgattag gctcagttac cggtcttttt
     6061 tttctcattt acactagcta agcatgctga taacttcgta tagcatacat tatacgaagt
     6121 tattaccctg ttatccctac aaattaaact gggcatcagc gcaattcaat tggtttggta
     6181 atagcaaggg aaaatagaat gaagtgatct ccaaaaaata agtacttttt gactgtaaat
     6241 aaaattgtaa ggagtaaaaa gtactttttt ttctaaaaaa atgtaattaa gtaaaagtaa
     6301 aagtattgat ttttaattgt actcaagtaa agtaaaaatc cccaaaaata atacttaagt
     6361 acagtaatca agtaaaatta ctcaagtact ttacacctct gggcccaatt cgccctatag
     6421 tgagtcgtat tacaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     6481 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     6541 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggacgcg
     6601 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6661 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6721 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6781 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6841 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6901 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6961 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     7021 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
     7081 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
     7141 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7201 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7261 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7321 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7381 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7441 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     7501 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7561 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7621 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     7681 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7741 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     7801 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7861 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7921 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7981 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     8041 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     8101 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     8161 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     8221 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     8281 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     8341 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8401 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     8461 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8521 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8581 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8641 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8701 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8761 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8821 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8881 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8941 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     9001 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     9061 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagc
//
