LOCUS Exported 7902 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCB168 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7902) AUTHORS Kiyomitsu T, Cheeseman IM TITLE Chromosome- and spindle-pole-derived signals generate an intrinsic code for spindle position and orientation. JOURNAL Nat Cell Biol. 2012 Feb 12;14(3):311-7. doi: 10.1038/ncb2440. PUBMED 22327364 REFERENCE 2 (bases 1 to 7902) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7902 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 8..477 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 542..741 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 934..1350 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1318..1334 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" regulatory 1385..1394 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1391..2107 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 1391..2107 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1436..1457) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1697..1716) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2044..2065 /label=EGFP-C /note="EGFP, forward primer" CDS 2153..2173 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 2186..2230 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" primer_bind complement(4317..4337) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4322..4651 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4502..4637 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4564..4583 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 4666..4713 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 4732..5130 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" LTR 5183..5776 /note="long terminal repeat from Moloney murine leukemia virus" primer_bind complement(5977..5994) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6148..6736) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6228..6247) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6914..7774) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 7537..7556 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7775..7879) /gene="bla" /label=AmpR promoter ORIGIN 1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca 61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt 121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag 181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc 241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga 301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc 361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc 421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt 481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg 541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta 601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa 661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc 721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg 781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt 841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt 901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt 961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc 1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg 1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca 1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct 1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt 1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt 1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc cagtgtggtg gtacgctacc 1381 ggtcgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt 1441 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga 1501 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc 1561 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga 1621 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg 1681 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg 1741 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat 1801 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa 1861 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt 1921 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc 1981 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga 2041 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct 2101 gtacaagtcc ggaggggatc ggtggagctc caccggtggc ggccgctcta gagagaatct 2161 ttattttcag ggcgccgcca aattcaaaga aaccgctgct gctaaattcg aacgccagca 2221 catggacagc ggaggtggag gtactagtct cgagatgtct gacaaaagtg acttaaaagc 2281 tgagctagag cgcaaaaagc agcgcttagc acagataaga gaagagaaga aacggaagga 2341 agaggagagg aaaaagaaag aggctgatat gcagcagaag aaagaacccg ttcaggacga 2401 ctctgatctg gatcgcaaac gacgagagac agaggctttg ctgcaaagca ttggtatctc 2461 accggagccg cctctagtcc caacccctat gtctccctcc tcgaaatcag tgagcactcc 2521 cagtgaagct ggaagccaag actcaggcga tctggggcca ttaacaagga ccctgcagtg 2581 ggacacagac ccctcagtgc tccagctgca gtcagactca gaacttggaa gaagactgca 2641 taaactgggc gtgtcaaagg tcacccaagt ggatttcctg ccaagggaag tagtgtccta 2701 ctcaaaggag acccagactc ctcttgccac gcatcagtct gaagaggatg aggaagatga 2761 ggaaatggtg gaatctaaag ttggccagga ctcagaactg gaaaatcagg acaaaaaaca 2821 ggaagtgaag gaagcccctc caagagagtt gacagaggaa gaaaaacagc agatcattca 2881 ttcagaggaa tttctcatct tttttgaccg gacaatacgg gtaattgaaa gagccctggc 2941 tgaagattcc gacatctttt ttgactacag cggccgagag ttagaggaaa aagatgggga 3001 tgttcaggct ggagccaatc tttctttcaa tcgtcagttc tatgatgaac attgttccaa 3061 gcatcgagtg gtcacttgta tggactggtc cctccagtac cctgagctga tggtggcttc 3121 ttacaacaac aatgaagatg ctccccatga accagatgga gtggccttgg tttggaacat 3181 gaagtttaag aaaaccacac cagaatacgt cttccactgt cagtcctctg tgatgtcggt 3241 ctgcttcgcc cgtttccatc ctaacttggt ggttggtggg acttactcgg gccagattgt 3301 cctctgggac aatcgcagtc atcgaaggac tccagtgcag cggacaccct tatcagctgc 3361 tgcacacacg catcccgtgt actgtgtaaa tgttgttggg acccagaatg ctcataacct 3421 catcactgtc tccactgatg gcaaaatgtg ttcctggagc ctggacatgc tctcaactcc 3481 acaggagagc atggagctgg tgtacaataa gtccaagcct gtcgctgtta ccggaatggc 3541 tttcccaacg ggagacgtca ataacttcgt ggttggcagt gaggaaggta cagtctacac 3601 ggcttgtcgt catggaagca aagcaggtat tggtgaggtc tttgaaggtc accaagggcc 3661 agtgacagga attaactgcc acatggcagt gggaccaatc gacttttctc acctgtttgt 3721 cacatcatca tttgactgga ctgtcaaact gtggaccacc aagcacaaca agccgctcta 3781 ctcctttgaa gacaatgcag actatgtgta cgatgtcatg tggtcccccg tgcatcctgc 3841 gctttttgcc tgcgtggacg ggatggggcg cttggacctc tggaacctca acaatgacac 3901 cgaggttcca acagcaagtg tggccattga gggggcatcc gccctaaacc gtgttcgttg 3961 ggcccaagct ggcaaagaag ttgctgttgg ggactcggaa ggccgtattt gggtctatga 4021 cgttggagag cttgcagttc cccacaatga tgaatggacc cgatttgcca ggacccttgt 4081 ggaaattcgt gctaacagag ctgatagcga ggaggaaggc actgttgagt tatctgccta 4141 ggaattctgc agtcgacggt accgcgggcc cgggatccac cggatctaga taactgatca 4201 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc 4261 ccctgaacct gaaacataaa atgaatgcaa ttcgccagca cagtggtcga ccctgtggaa 4321 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 4381 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 4441 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 4501 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 4561 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 4621 aggctttttt ggaggcctag gcttttgcaa aaagcttgag cacgtgttga caattaatca 4681 tcggcatagt atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag 4741 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc 4801 cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc 4861 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc 4921 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac 4981 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg 5041 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg 5101 ctgccctctg gttatgtgtg ggagggctaa gcacttcatc gataaaataa aagattttat 5161 ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt 5221 aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat 5281 caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca 5341 gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga 5401 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc 5461 ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc 5521 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc 5581 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc 5641 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat 5701 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca 5761 gcgggggtct ttcatttccg acttgtggtc tcgctgcctt gggagggtct cctctgagtg 5821 attgactacc cgtcagcggg ggtcttcaca tgcagcatgt atcaaaatta atttggtttt 5881 ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc caacgcgcgg 5941 ggagaggcgg tttgcgtatt ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 6001 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 6061 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 6121 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 6181 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 6241 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 6301 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 6361 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 6421 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 6481 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 6541 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 6601 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 6661 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 6721 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 6781 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 6841 ccttttaaat taaaaatgaa gtttgcggcc gcaaatcaat ctaaagtata tatgagtaaa 6901 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 6961 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 7021 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 7081 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 7141 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 7201 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 7261 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 7321 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 7381 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 7441 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 7501 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa 7561 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 7621 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 7681 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 7741 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 7801 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 7861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc ac //