LOCUS Exported 6670 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS FLM-PC1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6670) AUTHORS Talbot JJ, Shillingford JM, Vasanth S, Doerr N, Mukherjee S, Kinter MT, Watnick T, Weimbs T TITLE Polycystin-1 regulates STAT activity by a dual mechanism. JOURNAL Proc Natl Acad Sci U S A. 2011 May 10;108(19):7985-90. Epub 2011 Apr 25. PUBMED 21518865 REFERENCE 2 (bases 1 to 6670) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6670 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 41..59 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 62..80 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 90..114 /label=LNCX /note="Human CMV promoter, forward primer" CDS 1774..1803 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1813..1842 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS 1858..1875 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(1898..1915) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1904..2128 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2174..2602 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2261..2280) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2471..2492 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2611..2631) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2616..2946 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2797..2932 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2859..2878 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 2994..3041 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 3060..3434 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 3564..3685 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3601..3620) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3655..3674 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3734..3750) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3734..3750) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3747..3769) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3758..3774 /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(3782..3812) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3827..3848 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3965..3982) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4136..4724) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4216..4235) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4895..5755) /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 5518..5537 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5756..5860) /gene="bla" /label=AmpR promoter primer_bind complement(5935..5954) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 6126..6505 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 6506..39 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 6660..10 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" ORIGIN 1 ggtaggcgtg tacggtggga ggtctatata agcagagctc tccctatcag tgatagagat 61 ctccctatca gtgatagaga tcgtcgacga gctcgtttag tgaaccgtca gatcgcctgg 121 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg 181 actctagcgt ttaaacttaa gcttctagag atccctcgac ctcgagatcc attgtgctct 241 aaagatcttt ggtgacttgt ccactccagt gtggcatcat gtggcagctg ctcctcccaa 301 ctgctctgct acttctagtt tcagctggca tgcggactga agatctccca aaggctgtgg 361 tgttcctgga gcctcaatgg tacagcgtgc ttgagaagga cagtgtgact ctgaagtgcc 421 agggagccta ctcccctgag gacaattcca cacagtggtt tcacaatgag agcctcatct 481 caagccaggc ctcgagctac ttcattgacg ctgccacagt caacgacagt ggagagtaca 541 ggtgccagac aaacctctcc accctcagtg acccggtgca gctagaagtc catatcggct 601 ggctgttgct ccaggcccct cggtgggtgt tcaaggagga agaccctatt cacctgaggt 661 gtcacagctg gaagaacact gctctgcata aggtcacata tttacagaat ggcaaagaca 721 ggaagtattt tcatcataat tctgacttcc acattccaaa agccacactc aaagatagcg 781 gctcctactt ctgcaggggg cttgttggga gtaaaaatgt gtcttcagag actgtgaaca 841 tcaccatcac tcaaggtttg gcagtgtcaa ccatctcatc attctctcca cctggcgcgg 901 atccaccaag ggcctctgcc ctccctgccc caccgacagg ctccgccctc cctgacccgc 961 agacagcctc tgccctccct gacccgccag cagcctctgc cctccctgcg gccctggcgg 1021 tgatctcctt cctcctcggg ctgggcctgg gggtggcgtg tgtgctggcg aggacgcgtg 1081 agtcctggca cttgtcaccc ctgctgtgtg tggggctctg ggcactgcgg ctgtggggcg 1141 ccctacggct gggggctgtt attctccgct ggcgctacca cgccttgcgt ggagagctgt 1201 accggccggc ctgggagccc caggactacg agatggtgga gttgttcctg cgcaggctgc 1261 gcctctggat gggcctcagc aaggtcaagg agttccgcca caaagtccgc tttgaaggga 1321 tggagccgct gccctctcgc tcctccaggg gctccaaggt atccccggat gtgcccccac 1381 ccagcgctgg ctccgatgcc tcgcacccct ccacctcctc cagccagctg gatgggctga 1441 gcgtgagcct gggccggctg gggacaaggt gtgagcctga gccctcccgc ctccaagccg 1501 tgttcgaggc cctgctcacc cagtttgacc gactcaacca ggccacagag gacgtctacc 1561 agctggagca gcagctgcac agcctgcaag gccgcaggag cagccgggcg cccgccggat 1621 cttcccgtgg cccatccccg ggcctgcggc cagcactgcc cagccgcctt gcccgggcca 1681 gtcggggtgt ggacctggcc actggcccca gcaggacacc ccttcgggcc aagaacaagg 1741 tccaccccag cagcacttag ggcggccgct cgagaacaaa aactcatctc agaagaggat 1801 ctggggccct tcgaacaaaa actcatctca gaagaggatc tgaatatgca taccggtcat 1861 catcaccatc accattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc 1921 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc 1981 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct 2041 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg 2101 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct 2161 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2221 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 2281 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta 2341 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 2401 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 2461 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 2521 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt 2581 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 2641 ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2701 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat 2761 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt 2821 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc 2881 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt 2941 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca cgtgttgaca 3001 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca 3061 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg 3121 agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac ttcgccggtg 3181 tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca 3241 acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag tggtcggagg 3301 tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc ggcgagcagc 3361 cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg 3421 aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt 3481 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 3541 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 3601 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 3661 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 3721 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 3781 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 3841 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 3901 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 3961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4021 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 4141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 4201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 4261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 4321 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 4381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 4441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 4501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 4561 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 4621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 4681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 4741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 4801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 4861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 4921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 4981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 5101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 5221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 5281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 5341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 5401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 5461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 5521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 5581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 5641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 5701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 5761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 5821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 5881 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtcgactc tcagtacaat 5941 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc 6001 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat 6061 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac 6121 gcgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca 6181 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc 6241 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat 6301 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt 6361 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc 6421 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta 6481 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg 6541 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt 6601 gttttggaac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac 6661 gcaaatgggc //