LOCUS Exported 7900 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCFJ1209 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7900) TITLE Erik Jorgensen's plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 7900) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7900 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 283..299 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 283..299 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" protein_bind 319..339 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" promoter 340..939 /gene="eft-3" /label=eft-3 promoter /note="promoter for the C. elegans elongation factor 1-alpha gene (Mitrovich and Anderson, 2000)" protein_bind 965..989 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 992..1012 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1013..2437 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH LFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYE GTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMN FEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVL KGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYER SEGRHHLFLYGMDELYK" CDS 2447..2518 /codon_start=1 /product="nuclear localization signal from the C. elegans EGL-13 transcription factor (Lyssenko et al., 2007)" /label=EGL-13 NLS /translation="SRRRKANPTKLSENAKKLAKEVEN" protein_bind complement(2520..2544) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" terminator 2549..2872 /gene="tbb-2" /label=tbb-2 terminator /note="transcription terminator for the C. elegans tubulin beta-2 gene" protein_bind complement(2876..2896) /gene="mutant version of attB" /label=attB3 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" primer_bind complement(2904..2921) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2904..2920) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 3737..4531 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3791..3810) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4401..4420 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 5232..5248 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 5233..5249 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5785..5804) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5904..5926 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5964..5982) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6050..6154 /gene="bla" /label=AmpR promoter CDS 6155..7015 /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(6373..6392) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7186..7774 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7675..7694 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 gcaccgtacg tctcgaggaa ttctatcagg tgtacaagta tgaaatgtcg tttcgaacat 61 atagatgtct cgcaaacgta aatatttatc gattgtcata aaactttgac cttgtgaagt 121 gtcaaccttg actgtcgaac caccatagtt tggcgcgaat tgagcgtcat aattgtttac 181 tctcagtgca gtcaacatgt cgagtttcgt gccgaataaa gagcaaacgc ggacagtatt 241 aattttctgt tttcatttga agaaaacagc tgcggccgga tccaggaaac agctatgacc 301 atgattacgc caagctatca actttgtata gaaaagttgg cacctttggt cttttattgt 361 caacttccat tggttcttcc attgtttctg ttaaattaat gaatttttca taaaataaag 421 acattataca atataaaaat gaagaattta ttgaaaataa actgccagag agaaaaagta 481 tgcaacactc ccgccgagag tgtttgaaat ggtgtacggt acattttcgt gctaggagtt 541 agatgtgcag gcagcaacga gagggggaga gatttttttg ggccttgtga aattaacgtg 601 agttttctgg tcatctgact aatcatgttg gttttttgtt ggtttatttt gtttttatct 661 ttgtttttat ccagattagg aaatttaaat tttatgaatt tataatgagg tcaaacattc 721 agtcccagcg tttttcctgt tctcactgtt tagtcgaatt tttattttag gctttcaaca 781 aatgttctaa ctgtcttatt tgtgacctca ctttttatat ttttttaatt tttaaaaata 841 ttagaagttt ctaggataat tttttcgact tttattctct ctaccgtccg cactcttctt 901 acttttaaat taaattgttt ttttttcagt tgggaaacac tttgctcact ccgtagcagc 961 catggcaagt ttgtacaaaa aagcaggctc gccaaagaag aagcgtaagg tcgtgagcaa 1021 gggcgaggag gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa 1081 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac 1141 cgccaagctg aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc 1201 ccagttcatg tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa 1261 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg 1321 tctggtgacc gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa 1381 gatgcgcggc accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg 1441 ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca 1501 ggccctgaag ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc 1561 caagaagccc gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc 1621 ccacaacgag gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct 1681 gttcctgggg catggcaccg gcagcaccgg cagcggcagc tccggcaccg cctcctccga 1741 ggacaacaac atggccgtca tcaaagagtt catgcgcttc aaggtgcgca tggagggctc 1801 catgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac 1861 ccagaccgcc aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct 1921 gtccccccag ttcatgtacg gctccaaggc gtacgtgaag caccccgccg acatccccga 1981 ttacaagaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga 2041 cggcggtctg gtgaccgtga cccaggactc ctccctgcag gacggcacgc tgatctacaa 2101 ggtgaagatg cgcggcacca acttcccccc cgacggcccc gtaatgcaga agaagaccat 2161 gggctgggag gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagat 2221 ccaccaggcc ctgaagctga aggacggcgg ccactacctg gtggagttca agaccatcta 2281 catggccaag aagcccgtgc aactgcccgg ctactactac gtggacacca agctggacat 2341 cacctcccac aacgaggact acaccatcgt ggaacagtac gagcgctccg agggccgcca 2401 ccacctgttc ctgtacggca tggacgagct gtacaagtac ccagcttccc gtcgtcgtaa 2461 ggccaaccca accaagctct ccgagaacgc caagaagctc gccaaggagg tcgagaacaa 2521 cccagctttc ttgtacaaag tgggataaat gcaaaatcct ttcaagcatt cccttcttct 2581 ctatcactct tctttctttt tgtcaaaaaa ttctctcgct aatttatttg cttttttaat 2641 gttattattt tatgactttt tatagtcact gaaaagtttg catctgagtg aagtgaatgc 2701 tatcaaaatg tgattctgtc tgatgtactt tcacaatctc tcttcaattc cattttgaag 2761 tgctttaaac ccgaaaggtt gagaaaaatg cgagcgctca aatatttgta ttgtgttcgt 2821 tgagtgaccc aacaaaaaga ggaaacttta ttgtgccgcc aagaaaaaag tctcacaact 2881 ttattataca tagttgataa ttcactggcc gtcgttttac aggccggatg ggccatggca 2941 ccatttctaa attttagttt aataaaaaaa ttttaaaata atacgagttt tgagactaac 3001 tttcaataaa aaccaaaaaa ttgcttcaaa aaagagctcg cgccgattta cgataaacac 3061 agcgcgctac gacactccga aaaccgcgcc gcaccattac attctccgcc ccaccgcacg 3121 gtttgcggaa ggcgatgaaa ttgctgggcc taaaatttta accctttttt tcccgttttt 3181 caactgtttt cccgattttt tcgatatttt tttcaggttt ttagttagaa aagtatataa 3241 aaattgcatt atttgtagtt ttcgctcgta atttgtgata gaaaacggcc aatttgccta 3301 gatttgtgat tctaccaagt ggaatttttg ggaaaatatc aatattccac gatttttatt 3361 caaatcataa cgtttcgaaa ttatcagctt aaataaaact aaaataaata ataaaatact 3421 aaaaaccctg aatttctcgg ttctcggttc agcttgagtt cgttctatac acgttatcga 3481 tttttgtcat ttcgtttaat catgggtttt gattcatttt aaatatgaaa tttgatttat 3541 tcctcgtttc gaagttggtt tgtcttgtcc ggccggtaaa caaatgttat taaaaaaaat 3601 aataaatttc gaaatttttc gtaatgttcc gcaatcgtgt ttttccgttt atctcgtctt 3661 cccttccttc aaatatacta tttattttct caattcacat attttcagcc agccctgctc 3721 aacaagtgga ataaaaatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 3781 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 3841 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 3901 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt 3961 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 4021 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 4081 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 4141 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 4201 ggatgatctg gacgaagagc atcaggggct cgcaccagcc gaactgttcg ccaggctcaa 4261 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 4321 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 4381 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 4441 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 4501 cttctatcgc cttcttgacg agttcttctg agtccaatta ctcttcaaca tccctacatg 4561 ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact tctcttaatt 4621 tctttgtttt ttagcttctt ttaagccacc tctaacaatg aaattgtgta gattcaaaaa 4681 tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc attaataata 4741 attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt tacttctgat 4801 aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca tatgttacgt 4861 ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat gacgtcaaat 4921 catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa gtgaaagttt 4981 atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg tcaagatttc 5041 gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc aagaaagatc 5101 ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata atttgaaagt 5161 ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa tataccaaac 5221 ataactgttt ctcgaggtcg acggtatcga taagcttgat ccggatctgc ggcttactca 5281 ccagacctgg ccccatccga ttaccaccta ttcgcttcga tgggacacgc actcgctgag 5341 cagcgcttcg attcttacga aagtgtgaaa aaatggctcg atgaatggtt cgccgcaaaa 5401 gacgatgagt tctactggcg tggaatccac aaattgcccg agagatggga aaaatgtgta 5461 gctagcgacg gcaaatactt tgaataaatg attttttctt tttccacaaa atttaacgtg 5521 ttttttgatt taaaaaaaac gacatttcat acttgtacac ctgatatcta gatacctagg 5581 tgagctctgg tacccccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt 5641 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag 5701 cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc 5761 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc 5821 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc 5881 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc 5941 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat 6001 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc 6061 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg 6121 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc 6181 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt 6241 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct 6301 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac 6361 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact 6421 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa 6481 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga 6541 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt 6601 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga 6661 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg 6721 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat 6781 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat 6841 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc 6901 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga 6961 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc 7021 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag 7081 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc 7141 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt 7201 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt 7261 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat 7321 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc 7381 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa 7441 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg 7501 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag 7561 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag 7621 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa 7681 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt 7741 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg 7801 gttcctggtc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc 7861 tgtggataac cgtattaccg cctttgagtg agctgatacc //