LOCUS Exported 3488 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION human GFP11 with tagRFP 33bp downstream in Lamin A/C. ACCESSION . VERSION . KEYWORDS pAP1892-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3488) AUTHORS Paix A, Folkmann A, Goldman DH, Kulaga H, Grzelak MJ, Rasoloson D, Paidemarry S, Green R, Reed RR, Seydoux G TITLE Precision genome editing using synthesis-dependent repair of Cas9-induced DNA breaks. JOURNAL Proc Natl Acad Sci U S A. 2017 Nov 28. pii: 1711979114. doi: 10.1073/pnas.1711979114. PUBMED 29183983 REFERENCE 2 (bases 1 to 3488) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3488 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 105..124 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 318..340 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 332..349 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 333..349 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 376..423 /codon_start=1 /product="11th beta-strand of superfolder GFP" /label=GFP11 /note="associates with GFP(1-10), consisting of the first 10 beta-strands, to reconstitute fluorescent GFP (Cabantous et al., 2005; Kamiyama et al., 2016)" /translation="RDHMVLHEYVNAAGIT" primer_bind complement(1221..1237) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1221..1237) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1234..1256) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1245..1261 /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)." promoter complement(1269..1299) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1314..1335 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1452..1469) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1623..2211) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1703..1722) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2382..3242) /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 3005..3024 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3243..3347) /gene="bla" /label=AmpR promoter primer_bind 3415..3433 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3471..5) /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg 61 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat 121 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata 181 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg 241 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg 301 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct 361 cggtacccgg ggatccgtga ccacatggtc cttcatgagt atgtaaatgc tgctgggatt 421 acaggtggcg gccagcggcg cgccacccgc agcggggcgc aggccgtctc taaaggggaa 481 gaactcatca aggagaatat gcacatgaag ctctatatgg aaggtacagt aaacaatcac 541 cacttcaaat gcacttctga aggtgaagga aagccctacg agggtacaca gacgatgaga 601 attaaagtcg ttgagggagg cccgctgccc tttgcttttg acatccttgc aacctcattt 661 atgtatggca gccgaacatt catcaatcat acacagggga ttccagattt tttcaaacag 721 tcctttccgg agggctttac atgggagcga gtaacgacct atgaagatgg gggagttttg 781 accgccacgc aagatacgtc actccaagac ggctgcctga tttacaatgt aaaaataaga 841 ggcgttaatt ttccgtctaa tggcccggta atgcagaaaa agaccctggg ttgggaggcc 901 aacacggaaa tgctttatcc ggcagacggt ggcttggagg ggcgctctga catggcactg 961 aagttggtcg gcggaggtca ccttatttgc aacttcaaaa ctacgtaccg ctcaaagaag 1021 ccagccaaaa accttaaaat gcccggggtt tattatgtag accaccgcct cgaacggatc 1081 aaggaagctg ataaagaaac gtatgtagag caacatgaag tcgcagtggc aaggtactgt 1141 gatctcccca gtaagctggg gcacaaactg aatgatcctc tagagtcgac ctgcaggcat 1201 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 1261 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 1321 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 1381 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 1441 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 1501 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 1561 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 1621 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 1681 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 1741 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 1801 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 1861 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 1921 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 1981 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 2041 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 2101 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 2161 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 2221 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 2281 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 2341 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 2401 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 2461 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 2521 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 2581 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 2641 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 2701 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 2761 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 2821 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 2881 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 2941 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 3001 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 3061 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 3121 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 3181 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 3241 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 3301 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 3361 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 3421 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 3481 atgcagct //