LOCUS Exported 9119 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGBT-RP2-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9119) AUTHORS Clark KJ, Balciunas D, Pogoda HM, Ding Y, Westcot SE, Bedell VM, Greenwood TM, Urban MD, Skuster KJ, Petzold AM, Ni J, Nielsen AL, Patowary A, Scaria V, Sivasubbu S, Xu X, Hammerschmidt M, Ekker SC TITLE In vivo protein trapping produces a functional expression codex of the vertebrate proteome. JOURNAL Nat Methods. 2011 Jun;8(6):506-15. Epub 2011 May 8. PUBMED 21552255 REFERENCE 2 (bases 1 to 9119) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9119 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 22..43 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /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 /bound_moiety="lac repressor encoded by lacI" /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 101..123 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 120..136 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 120..136 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 154..172 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 154..171 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 448..481 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 653..1327 /codon_start=1 /product="monomeric derivative of DsRed (Campbell et al., 2002)" /label=mRFP1 /note="mammalian codon-optimized" /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGVV TVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKMR LKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHST GA" primer_bind complement(780..798) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1048..1067 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1242..1265 /label=DsRed1-C /note="DsRed1, forward primer" primer_bind 2575..2595 /label=T3 /note="T3 promoter, forward primer" promoter 2577..2595 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 5165..5881 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MSKGEELFTGVVPILVELNGDVNGHKFPVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind 5818..5838 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" protein_bind 6090..6123 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(6413..6432) /label=T7 /note="T7 promoter, forward primer" promoter complement(6414..6432) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6439..6456) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6439..6455) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6448..6470) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6596..7051 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6683..6702) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6893..6914 /label=F1ori-F /note="F1 origin, forward primer" promoter 7129..7233 /gene="bla" /label=AmpR promoter CDS 7234..8094 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(7452..7471) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8265..8853 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8754..8773 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9007..9024 /label=L4440 /note="L4440 vector, forward primer" 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 //