LOCUS Exported 7953 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDZ306 (GLT1 POL1 24xPP7SL) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7953) AUTHORS Larson DR, Zenklusen D, Wu B, Chao JA, Singer RH TITLE Real-time observation of transcription initiation and elongation on an endogenous yeast gene. JOURNAL Science. 2011 Apr 22;332(6028):475-8. PUBMED 21512033 REFERENCE 2 (bases 1 to 7953) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7953 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 416..759 /label=TEF promoter /note="Ashbya gossypii TEF promoter" terminator 1462..1659 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(4028..4047) /label=T7 /note="T7 promoter, forward primer" promoter complement(4029..4047) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4054..4071) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(4054..4070) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4063..4085) /label=M13/pUC Forward /note="In lacZ gene" CDS 4208..4510 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 4389..4408 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 4859..5653 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4913..4932) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5523..5542 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS complement(6199..6577) /label=ISS /note="immunostimulatory sequence from the AmpR gene; contains unmethylated CpG dinucleotides in the context of 5'-AACGTT-3' (Sato et al., 1996)" primer_bind 6524..6543 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6885..7473 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7374..7393 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7627..7644 /label=L4440 /note="L4440 vector, forward primer" protein_bind 7761..7782 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 7797..7827 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7835..7851 /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 7840..7862 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 7859..7875 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 7859..7875 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 7896..7914 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 ttaattaaac cgttcaaccc tatacagcgc aaatttggtc ctaatacact tcggttttaa 61 tgcgtcaatc cgattggctc cgataagctt ttgcacattt ttcaagtcat atgtcacgac 121 gaacgtgatg cccgcaaaac gtaaaaaaaa gaagcaataa attgcccaaa aaaaaaatat 181 aaactgcttt tcttagtcat tcataagttc tgatggtctt ccttatctta ttgtattttt 241 cgtcctattg tttcatttct taccaaatta atcaattctt atatcttact tgataacaca 301 ccaaactaat cgtctccaca tcataggaag ataggaaatt gctatctcag tcctatacta 361 cgcagacctc gagtctgttt agcttgcctc gtccccgccg ggtcacccgg ccagcgacat 421 ggaggcccag aataccctcc ttgacagtct tgacgtgcgc agctcagggg catgatgtga 481 ctgtcgcccg tacatttagc ccatacatcc ccatgtataa tcatttgcat ccatacattt 541 tgatggccgc acggcgcgaa gcaaaaatta cggctcctcg ctgcagacct gcgagcaggg 601 aaacgctccc ctcacagacg cgttgaattg tccccacgcc gcgcccctgt agagaaatat 661 aaaaggttag gatttgccac tgaggttctt ctttcatata cttcctttta aaatcttgct 721 aggatacagt tctcacatca catccgaaca taaacaacca tggcagaacc agcccaaaaa 781 aagcaaaaac aaactgttca ggagcgcaag gcgtttatct cccgtatcac taatgaaact 841 aaaattcaaa tcgctatttc gctgaatggt ggttatattc aaataaaaga ttcgattctt 901 cctgcaaaga aggatgacga tgtagcttcc caagctactc agtcacaggt catcgatatt 961 cacacaggtg ttggcttttt ggatcatatg atccatgcgt tggcaaaaca ctctggttgg 1021 tctcttattg ttgaatgtat tggtgacctg cacattgacg atcaccatac taccgaagat 1081 tgcggtatcg cattagggca agcgttcaaa gaagcaatgg gtgctgtccg tggtgtaaaa 1141 agattcggta ctgggttcgc accattggat gaggcgctat cacgtgccgt agtcgattta 1201 tctagtagac catttgctgt aatcgacctt ggattgaaga gagagatgat tggtgattta 1261 tccactgaaa tgattccaca ctttttggaa agtttcgcgg aggcggccag aattactttg 1321 catgttgatt gtctgagagg tttcaacgat caccacagaa gtgagagtgc gttcaaggct 1381 ttggctgttg ccataagaga agctatttct agcaatggca ccaatgacgt tccctcaacc 1441 aaaggtgttt tgatgtgaag tactgacaat aaaaagattc ttgttttcaa gaacttgtca 1501 tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag cgtgatttat 1561 attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca gaaagtaata 1621 tcatgcgtca atcgtatgtg aatgctggtc gctatactgc tgtcgattcg atactaacgc 1681 cgccatccag agctctacat gaactgaccg aaattgcagc atccttacaa aaacggatac 1741 atacgttaga aacggaaaac aagcttttaa agaatttagt tctgagtagc ggtgaaacgg 1801 aaggaataaa aaaagctgaa agcttaaaga agcaaatttt tgagaaggtt cagaaagaat 1861 aaaataacag actatacagt ttaattatat atatatatat atatacgtgc atttttctta 1921 aagaaatata acttttttct tgtactgcct gcaatctcta ttcttcattc atcacacatc 1981 tattcaaacg cgttaaaatt tttttgttag cctataagta aaacgcgtcg cgtgtatcca 2041 attttactga attaaattct ttactcattg gatgatatat atcttcttgg tctagacaac 2101 gaacacatac caattcagac tcacgagaaa aaaaactgtc ttcactctgc tattagaaac 2161 caaatagtat caaggcataa aataaacagt ggaaaggatc ctaaggtacc taattgccta 2221 gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac cagcagagca 2281 tatgggctcg ctggctgcag tattcccggg ttcattagat cctaaggtac ctaattgcct 2341 agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa ccagcagagc 2401 atatgggctc gctggctgca gtattcccgg gttcattaga tcctaaggta cctaattgcc 2461 tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc gactctagaa accagcagag 2521 catatgggct cgctggctgc agtattcccg ggttcattag atcctaaggt acctaattgc 2581 ctagaaagga gcagacgata tggcgtcgct ccctgcaggt cgactctaga aaccagcaga 2641 gcatatgggc tcgctggctg cagtattccc gggttcatta gatcctaagg tacctaattg 2701 cctagaaagg agcagacgat atggcgtcgc tccctgcagg tcgactctag aaaccagcag 2761 agcatatggg ctcgctggct gcagtattcc cgggttcatt agatcctaag gtacctaatt 2821 gcctagaaag gagcagacga tatggcgtcg ctccctgcag gtcgactcta gaaaccagca 2881 gagcatatgg gctcgctggc tgcagtattc ccgggttcat tagatcctaa ggtacctaat 2941 tgcctagaaa ggagcagacg atatggcgtc gctccctgca ggtcgactct agaaaccagc 3001 agagcatatg ggctcgctgg ctgcagtatt cccgggttca ttagatccta aggtacctaa 3061 ttgcctagaa aggagcagac gatatggcgt cgctccctgc aggtcgactc tagaaaccag 3121 cagagcatat gggctcgctg gctgcagtat tcccgggttc attagatcct aaggtaccta 3181 attgcctaga aaggagcaga cgatatggcg tcgctccctg caggtcgact ctagaaacca 3241 gcagagcata tgggctcgct ggctgcagta ttcccgggtt cattagatcc taaggtacct 3301 aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac tctagaaacc 3361 agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc ctaaggtacc 3421 taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga ctctagaaac 3481 cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat cctaaggtac 3541 ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg actctagaaa 3601 ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga tccatgtccc 3661 aggatagaat tttgttagat agatctatgc cagtgttgaa atcagacaat ttcgatccat 3721 tggaagaagc ttacgaaggt gggacaattc aaaactataa cgatgaacac catcttcata 3781 aatcttgggc aaatgtgatt ccggacaaac gaggacttta cgaccctgat tatgaacatg 3841 acgcttgtgg tgtcggtttc gtagcaaata agcatggtga acagtctcac aagattgtta 3901 ctgacgctag atatctttta gtgaatatga cacatcgtgg tgccgtctca tctgatggga 3961 acggtgacgg tgccggtatt ctgctaggta tcccaagggc gaattcgcgg ccgctaaatt 4021 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga 4081 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 4141 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctata 4201 cgtacggcag tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga 4261 tgtacagagt gatattattg acacgccggg gcgacggatg gtgatccccc tggccagtgc 4321 acgtctgctg tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga 4381 aagctggcgc atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga 4441 agtggctgat ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg 4501 gggaatataa atgtcaggca tgagattatc aaaaaggatc ttcacctaga tccttttcac 4561 gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat 4621 ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg 4681 gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc 4741 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag 4801 gatctgatgg cgcaggggat caagctctga tcaagagaca ggatgaggat cgtttcgcat 4861 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg 4921 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc 4981 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca 5041 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct 5101 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga 5161 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg 5221 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat 5281 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga 5341 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg 5401 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg 5461 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat 5521 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct 5581 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga 5641 cgagttcttc tgaattatta acgcttacaa tttcctgatg cggtattttc tccttacgca 5701 tctgtgcggt atttcacacc gcatcaggtg gcacttttcg gggaaatgtg cgcggaaccc 5761 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga ttatcaaaaa 5821 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat 5881 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga 5941 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac 6001 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccac gctcaccggc 6061 tccagattta tcagcaataa accagccagc cggaaggccg agcgcagaag tggtcctgca 6121 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 6181 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 6241 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 6301 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 6361 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6421 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6481 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6541 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6601 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6661 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 6721 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 6781 tattgaagca tttatcaggg ttattgtctc atgaccaaaa tcccttaacg tgagttttcg 6841 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 6901 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 6961 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 7021 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 7081 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 7141 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 7201 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 7261 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 7321 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 7381 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 7441 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 7501 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 7561 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc 7621 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc 7681 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg 7741 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta 7801 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca 7861 ggaaacagct atgaccatga ttacgccaag ctcagaatta accctcacta aagggactag 7921 tcctgcaggt ttaaacgaat tcgcccttat ata //