LOCUS Exported 5622 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGBT-R16 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5622) 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 5622) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5622 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind complement(221..254) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS 581..1255 /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(708..726) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 976..995 /label=mCherry-F /note="mCherry, forward primer" primer_bind 1170..1193 /label=DsRed1-C /note="DsRed1, forward primer" protein_bind complement(2366..2399) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind complement(2695..2714) /label=T7 /note="T7 promoter, forward primer" promoter complement(2696..2714) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2721..2738) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2721..2737) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2730..2752) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2878..3333 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2965..2984) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3175..3196 /label=F1ori-F /note="F1 origin, forward primer" promoter 3411..3515 /gene="bla" /label=AmpR promoter CDS 3516..4376 /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(3734..3753) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4547..5135 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5036..5055 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5289..5306 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5423..5444 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5459..5489 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5497..5513 /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 5502..5524 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5521..5537 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5521..5537 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 5555..5573 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 5555..5572 /label=SP6 /note="SP6 promoter, forward primer" 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 gctgtaggga taacagggta ataacttcgt ataatgtatg 241 ctatacgaag ttatcagcat gcaaaccgtt ctctgaagcc tctcgtctgt cccgtttcct 301 cattttaagt tctcctgttc attggttcac ttcctccagg ctttgtgtcc tctgagctcc 361 tgagtttctc ctcttctcag caggttatct aaactttgcc tgcctgtttt gcagttcctt 421 ctgcactttt tgttccttta ctctggattt ccgactaacc cctgcatgga agtgtggatt 481 gtgtgctgca gacacttgaa catccagtta acgtctccat cctctctttg cagatcatgt 541 tggagacctt caacaccccc gcccccgggt accggaaggt gcctcctccg aggacgtcat 601 caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc acgagttcga 661 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt 721 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tccagtacgg 781 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc 841 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac 901 ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa 961 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga 1021 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagatgaggc tgaagctgaa 1081 ggacggcggc cactacgacg ccgaggtcaa gaccacctac atggccaaga agcccgtgca 1141 gctgcccggc gcctacaaga ccgacatcaa gctggacatc acctcccaca acgaggacta 1201 caccatcgtg gaacagtacg agcgcgccga gggccgccac tccaccggcg cctaagaaat 1261 cgatacggac tgttaccact tcacgccgac tcaaactgcg cagagaaact tcaaacgaca 1321 acattggcat ggcttttgtt atttttggcg cttgactcag gatctaaaaa ctggaacggt 1381 gaaggtgacg gcaatgtttt ttggcaaata agcatccccg aagttctaca atgcatctga 1441 ggactccatg ttttgttttc tttagtcatt ccaaatgttt gttaaatgca ttgttccgaa 1501 acttatttgc ctctatgaag gctgcccagt aattgggagc atacttaaca ttgtagtatt 1561 gtatgtaaat tatgtaacaa aacaatgtct gggtttttgt acttttcagc cttaaaatct 1621 tgggtttatt tttttttctt tgttccaaaa aaacgaagct ttaccattca agatgtaaag 1681 gtttccattc cccctgggca tattgtaaaa gctgtgtgga acgtggcggt gccagacatt 1741 tggtggggcc aacctgtaca ctgactaatt caattccaat aaaagtgcac atgtaagaca 1801 tcctacccct gtctcatttc ctgtcttggc tctgagtggc ttcaataaaa atggcttttg 1861 tgaagttgca ccatgttaat tacatttcag cactttcaca ccaggttttt gactaatttg 1921 aatttatcga agtttgacct tttttttttt ttttccttca agctaatttg aggttatcaa 1981 accatttcat tggaggaaac ttaatgttta aaagcataaa ttcgaaacat gattcattta 2041 tttaactgct taatggtgaa acaaacttca agtaatacac caagtgttcc ctcatatgaa 2101 ggtagctaac tgcaaacggc cttaactttc catcttaagt tgagctacat aaaggcttta 2161 acaggaagga agagccagtt taaaagtcaa accaccatga ctgatataaa ttcttgtcca 2221 tataaccttg cgtttcacac aatatagcta tctactgcta aataaattat cgcgattcct 2281 attctctaga aagtatagga acttcctagg agcgctccgg agatcttcct attctctaga 2341 aagtatagga acttcttaat taacgataac ttcgtataat gtatgctata cgaagttata 2401 ttaccctgtt atccctacca tgaaattaaa ctgggcatca gcgcaattca attggtttgg 2461 taatagcaag ggaaaataga atgaagtgat ctccaaaaaa taagtacttt ttgactgtaa 2521 ataaaattgt aaggagtaaa aagtactttt ttttctaaaa aaatgtaatt aagtaaaagt 2581 aaaagtattg atttttaatt gtactcaagt aaagtaaaaa tccccaaaaa taatacttaa 2641 gtacagtaat caagtaaaat tactcaagta ctttacacct ctgggcccaa ttcgccctat 2701 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct 2761 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc 2821 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg 2881 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 2941 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 3001 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 3061 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 3121 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 3181 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 3241 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 3301 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca 3361 tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc 3421 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct 3481 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg 3541 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg 3601 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc 3661 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca 3721 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac 3781 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa 3841 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg 3901 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt 3961 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg 4021 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc 4081 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga 4141 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta 4201 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc 4261 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg 4321 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt 4381 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa 4441 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt 4501 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt 4561 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt 4621 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga 4681 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag 4741 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata 4801 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg 4861 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga 4921 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca 4981 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa 5041 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt 5101 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac 5161 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt 5221 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga 5281 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc 5341 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag 5401 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt 5461 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca 5521 caggaaacag ctatgaccat gattacgcca agctatttag gtgacactat agaatactca 5581 agctatgcat ccaacgcgtt gggagctctc ccatatggtc ga //