LOCUS Exported 7510 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS TAp63alpha-FLAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7510) TITLE Sidransky Plasmids REFERENCE 2 (bases 1 to 7510) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7510 /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" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2858..2881 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(2932..2949) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2938..3162 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3208..3636 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3295..3314) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3505..3526 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3645..3665) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3650..3979 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3830..3965 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3892..3911 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4028..5053 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 5183..5304 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5220..5239) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5274..5293 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5353..5369) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5353..5369) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5366..5388) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5377..5393 /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(5401..5431) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5446..5467 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5584..5601) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5755..6343) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5835..5854) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6514..7374) /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 7137..7156 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7375..7479) /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 gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccatgtcc cagagcacac agacaaatga 961 attcctcagt ccagaggttt tccagcatat ctgggatttt ctggaacagc ctatatgttc 1021 agttcagccc attgacttga actttgtgga tgaaccatca gaagatggtg cgacaaacaa 1081 gattgagatt agcatggact gtatccgcat gcaggactcg gacctgagtg accccatgtg 1141 gccacagtac acgaacctgg ggctcctgaa cagcatggac cagcagattc agaacggctc 1201 ctcgtccacc agtccctata acacagacca cgcgcagaac agcgtcacgg cgccctcgcc 1261 ctacgcacag cccagctcca ccttcgatgc tctctctcca tcacccgcca tcccctccaa 1321 caccgactac ccaggcccgc acagtttcga cgtgtccttc cagcagtcga gcaccgccaa 1381 gtcggccacc tggacgtatt ccactgaact gaagaaactc tactgccaaa ttgcaaagac 1441 atgccccatc cagatcaagg tgatgacccc acctcctcag ggagctgtta tccgcgccat 1501 gcctgtctac aaaaaagctg agcacgtcac ggaggtggtg aagcggtgcc ccaaccatga 1561 gctgagccgt gaattcaacg agggacagat tgcccctcct agtcatttga ttcgagtaga 1621 ggggaacagc catgcccagt atgtagaaga tcccatcaca ggaagacaga gtgtgctggt 1681 accttatgag ccaccccagg ttggcactga attcacgaca gtcttgtaca atttcatgtg 1741 taacagcagt tgtgttggag ggatgaaccg ccgtccaatt ttaatcattg ttactctgga 1801 aaccagagat gggcaagtcc tgggccgacg ctgctttgag gcccggatct gtgcttgccc 1861 aggaagagac aggaaggcgg atgaagatag catcagaaag cagcaagttt cggacagtac 1921 aaagaacggt gatggcacga agcgcccgtt tcgtcagaac acacatggta tccagatgac 1981 atccatcaag aaacgaagat ccccagatga tgaactgtta tacttaccag tgaggggccg 2041 tgagacttat gaaatgctgt tgaagatcaa agagtccctg gaactcatgc agtaccttcc 2101 tcagcacaca attgaaacgt acaggcaaca gcaacagcag cagcaccagc acttacttca 2161 gaaacagacc tcaatacagt ctccatcttc atatggtaac agctccccac ctctgaacaa 2221 aatgaacagc atgaacaagc tgccttctgt gagccagctt atcaaccctc agcagcgcaa 2281 cgccctcact cctacaacca ttcctgatgg catgggagcc aacattccca tgatgggcac 2341 ccacatgcca atggctggag acatgaatgg actcagcccc acccaggcac tccctccccc 2401 actctccatg ccatccacct cccactgcac acccccacct ccgtatccca cagattgcag 2461 cattgtcagt ttcttagcga ggttgggctg ttcatcatgt ctggactatt tcacgaccca 2521 ggggctgacc accatctatc agattgagca ttactccatg gatgatctgg caagtctgaa 2581 aatccctgag caatttcgac atgcgatctg gaagggcatc ctggaccacc ggcagctcca 2641 cgaattctcc tccccttctc atctcctgcg gaccccaagc agtgcctcta cagtcagtgt 2701 gggctccagt gagacccggg gtgagcgtgt tattgatgct gtgcgattca ccctccgcca 2761 gaccatctct ttcccacccc gagatgagtg gaatgacttc aactttgaca tggatgctcg 2821 ccgcaataag caacagcgca tcaaagagga gggggaggac tacaaggacg acgatgacaa 2881 gtagtaggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca gcctcgactg 2941 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 3001 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 3061 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 3121 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa 3181 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg 3241 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 3301 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 3361 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 3421 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 3481 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 3541 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 3601 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt 3661 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 3721 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 3781 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct 3841 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc 3901 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg 3961 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca 4021 gcacgtgatg aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa 4081 gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag 4141 cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta 4201 caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct 4261 tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt 4321 cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc 4381 catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc 4441 gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca 4501 tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct 4561 cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga 4621 tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag 4681 cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg 4741 gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg 4801 atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt 4861 ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg 4921 atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac 4981 cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag 5041 ggcaaaggaa tagcacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt 5101 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 5161 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 5221 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 5281 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 5341 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 5401 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 5461 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 5521 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 5581 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 5641 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 5701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 5761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 5821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5941 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 6241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt 6301 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt 6361 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag 6421 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat 6481 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 6541 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 6601 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 6661 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 6721 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 6781 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 6841 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 6901 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 6961 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 7021 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 7081 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 7141 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 7201 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 7261 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 7321 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 7381 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 7441 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc 7501 acctgacgtc //