LOCUS Exported 7042 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3 Hygro HA Akt2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7042) TITLE Birnbaum plasmids REFERENCE 2 (bases 1 to 7042) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7042 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..820 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 771..791 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 865..884 /label=T7 /note="T7 promoter, forward primer" promoter 865..883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 930..956 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(2466..2483) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2472..2696 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2742..3170 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2829..2848) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3039..3060 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3179..3199) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3184..3513 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3364..3499 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3426..3445 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3562..4587 /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 4717..4838 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4754..4773) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4808..4827 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4887..4903) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4887..4903) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4900..4922) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4911..4927 /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(4935..4965) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4980..5001 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5118..5135) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5289..5877) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5369..5388) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6048..6908) /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 6671..6690 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6909..7013) /gene="bla" /label=AmpR promoter ORIGIN 1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga 61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct 181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca 241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata 301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc 841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta 901 gcgtttaaac ttaagcttgc gccaccatgt acccttatga tgtgccagat tatgccaatg 961 aggtgtctgt catcaaagaa ggctggctcc acaagcgtgg tgaatacatc aagacctgga 1021 ggccacggta cttcctgctg aagagcgacg gctccttcat tgggtacaag gagaggcccg 1081 aggcccctga tcagactcta ccccccttaa acaacttctc cgtagcagaa tgccagctga 1141 tgaagaccga gaggccgcga cccaacacct ttgtcatacg ctgcctgcag tggaccacag 1201 tcatcgagag gaccttccac gtggattctc cagacgagag ggaggagtgg atgcgggcca 1261 tccagatggt cgccaacagc ctcaagcagc gggccccagg cgaggacccc atggactaca 1321 agtgtggctc ccccagtgac tcctccacga ctgaggagat ggaagtggcg gtcagcaagg 1381 cacgggctaa agtgaccatg aatgacttcg actatctcaa actccttggc aagggaacct 1441 ttggcaaagt catcctggtg cgggagaagg ccactggccg ctactacgcc atgaagatcc 1501 tgcggaagga agtcatcatt gccaaggatg aagtcgctca cacagtcacc gagagccggg 1561 tcctccagaa caccaggcac ccgttcctca ctgcgctgaa gtatgccttc cagacccacg 1621 accgcctgtg ctttgtgatg gagtatgcca acgggggtga gctgttcttc cacctgtccc 1681 gggagcgtgt cttcacagag gagcgggccc ggttttatgg tgcagagatt gtctcggctc 1741 ttgagtactt gcactcgcgg gacgtggtat accgcgacat caagctggaa aacctcatgc 1801 tggacaaaga tggccacatc aagatcactg actttggcct ctgcaaagag ggcatcagtg 1861 acggggccac catgaaaacc ttctgtggga ccccggagta cctggcgcct gaggtgctgg 1921 aggacaatga ctatggccgg gccgtggact ggtgggggct gggtgtggtc atgtacgaga 1981 tgatgtgcgg ccgcctgccc ttctacaacc aggaccacga gcgcctcttc gagctcatcc 2041 tcatggaaga gatccgcttc ccgcgcacgc tcagccccga ggccaagtcc ctgcttgctg 2101 ggctgcttaa gaaggacccc aagcagaggc ttggtggggg gcccagcgat gccaaggagg 2161 tcatggagca caggttcttc ctcagcatca actggcagga cgtggtccag aagaagctcc 2221 tgccaccctt caaacctcag gtcacgtccg aggtcgacac aaggtacttc gatgatgaat 2281 ttaccgccca gtccatcaca atcacacccc ctgaccgcta tgacagcctg ggcttactgg 2341 agctggacca gcggacccac ttcccccagt tctcctactc ggccagcatc cgcgagtgaa 2401 gatctaggat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg 2461 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc 2521 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc 2581 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa 2641 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc 2701 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc 2761 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct 2821 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg 2881 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga 2941 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt 3001 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg 3061 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc 3121 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg 3181 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa 3241 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc 3301 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg 3361 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt 3421 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga 3481 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt 3541 ttcggatctg atcagcacgt gatgaaaaag cctgaactca ccgcgacgtc tgtcgagaag 3601 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa 3661 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc 3721 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg 3781 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc 3841 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag 3901 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc 3961 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg 4021 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc 4081 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac 4141 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg 4201 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc 4261 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat 4321 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa 4381 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc 4441 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc 4501 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc 4561 agcactcgtc cgagggcaaa ggaatagcac gtgctacgag atttcgattc caccgccgcc 4621 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag 4681 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat 4741 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 4801 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc 4861 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg 4921 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa 4981 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac 5041 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt 5101 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga 5161 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca 5221 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg 5281 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 5341 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 5401 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 5461 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 5521 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 5581 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 5641 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 5701 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag 5761 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 5821 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 5881 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 5941 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga 6001 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta 6061 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc 6121 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg 6181 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga 6241 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt 6301 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt 6361 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc 6421 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc 6481 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca 6541 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag 6601 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg 6661 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa 6721 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa 6781 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga 6841 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga 6901 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg 6961 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt 7021 ccccgaaaag tgccacctga cg //