LOCUS Exported 6823 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDN-D2irC6kwh SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6823) AUTHORS Nevozhay D, Zal T, Balazsi G TITLE Transferring a synthetic gene circuit from yeast to mammalian cells. JOURNAL Nat Commun. 2013 Feb 5;4:1451. doi: 10.1038/ncomms2471. PUBMED 23385595 REFERENCE 2 (bases 1 to 6823) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6823 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 814..832 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 840..858 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" intron 1041..1613 /label=-beta-globin intron /note="intron from rabbit beta-globin gene" regulatory 1625..1634 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1631..2341 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(1776..1794) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2044..2063 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(2409..2429) /label=WPRE-R /note="WPRE, reverse primer" primer_bind complement(2782..2799) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2788..3012 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3058..3486 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3145..3164) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3355..3376 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3495..3515) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3500..3829 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3680..3815 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3742..3761 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 3877..3924 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 3943..4341 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 4499..4620 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4536..4555) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4590..4609 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4669..4685) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4669..4685) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4682..4704) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4693..4709 /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(4717..4747) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4762..4783 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4900..4917) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5071..5656) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5151..5170) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5827..6687) /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 6450..6469 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6688..6792) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcatccctat cagtgataga gatcagatct 841 ccctatcagt gatagagagc tgttttgacc tccatagaag acaccgggac cgatccagcc 901 tccggactct agcgtttaaa cttaagcttg gtacccgggg atcctctagg gcctctgagc 961 tattccagaa gtagtgaaga ggcttttttg gaggcctagg cttttgcaaa aagctccgga 1021 tcgatcctga gaacttcagg gtgagtttgg ggacccttga ttgttctttc tttttcgcta 1081 ttgtaaaatt catgttatat ggagggggca aagttttcag ggtgttgttt agaatgggaa 1141 gatgtccctt gtatcaccat ggaccctcat gataattttg tttctttcac tttctactct 1201 gttgacaacc attgtctcct cttattttct tttcattttc tgtaactttt tcgttaaact 1261 ttagcttgca tttgtaacga atttttaaat tcacttttgt ttatttgtca gattgtaagt 1321 actttctcta atcacttttt tttcaaggca atcagggtat attatattgt acttcagcac 1381 agttttagag aacaattgtt ataattaaat gataaggtag aatatttctg catataaatt 1441 ctggctggcg tggaaatatt cttattggta gaaacaacta catcctggtc atcatcctgc 1501 ctttctcttt atggttacaa tgatatacac tgtttgagat gaggataaaa tactctgagt 1561 ccaaaccggg cccctctgct aaccatgttc atgccttctt ctttttccta caggtcctgc 1621 aggcgccacc atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat 1681 gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga 1741 gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg 1801 ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta 1861 cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa 1921 gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc 1981 cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga 2041 cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc 2101 cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca 2161 ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc 2221 ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga 2281 acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta 2341 gaccggtgcg gccgcaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 2401 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 2461 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 2521 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 2581 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 2641 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 2701 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcctcgag tctagagggc 2761 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 2821 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 2881 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 2941 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 3001 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 3061 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 3121 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 3181 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg 3241 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 3301 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 3361 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 3421 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 3481 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc 3541 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc 3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt 3661 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc 3721 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct 3781 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg 3841 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag 3901 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatggccaa gcctttgtct 3961 caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat ccccatctct 4021 gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt cactggtgtc 4081 aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg cactgctgct 4141 gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc 4201 ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc 4261 atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt gctgccctct 4321 ggttatgtgt gggagggcta agcacttcgt ggccgaggag caggactgac acgtgctacg 4381 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga 4441 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa 4501 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 4561 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 4621 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt 4681 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa 4741 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact 4801 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc 4861 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 4921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 4981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 5041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 5101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 5161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 5221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag 5281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 5341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 5401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 5461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt 5521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 5581 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag 5641 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 5701 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 5761 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 5821 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 5881 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 5941 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 6001 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 6061 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 6121 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 6181 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 6241 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 6301 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 6361 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 6421 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 6481 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 6541 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 6601 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 6661 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 6721 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 6781 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc //