LOCUS 40924_39143 5745 bp DNA circular SYN 13-DEC-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5745) AUTHORS caoheibi TITLE Direct Submission REFERENCE 2 (bases 1 to 5745) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..5745 /mol_type="other DNA" /organism="synthetic DNA construct" 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" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" sig_peptide 905..967 /label=Ig-kappa leader /note="leader sequence from mouse immunoglobulin kappa light chain" CDS 1082..1111 /codon_start=1 /label=Myc /note="Myc (human c-Myc proto-oncogene) epitope tag" /translation="EQKLISEEDL" CDS 1127..1144 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHHH" polyA_signal 1173..1397 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1443..1871 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 1885..2214 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2263..3285 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 3418..3551 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3588..3604) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(3612..3628) /label=lac operator /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(3636..3666) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(3681..3702) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(3990..4578) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4752..5609) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(5610..5714) /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 caccatggag acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac 961 tggtgacgcg gcccagccgg ccaggcgcgc cgtacgaagc ttggtaccga gctcggatcc 1021 actccagtgt ggtggaattc tgcagatatc cagcacagtg gcggccgctc gaggagggcc 1081 cgaacaaaaa ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcatcatca 1141 tcattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt 1201 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc 1261 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg 1321 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga 1381 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc 1441 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac 1501 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc 1561 cacgttcgcc ggctttcccc gtcaagctct aaatcggggc atccctttag ggttccgatt 1621 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg 1681 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag 1741 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt 1801 ataagggatt ttggggattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt 1861 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc 1921 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag 1981 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc 2041 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct 2101 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct 2161 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc 2221 ccgggagctt gtatatccat tttcggatct gatcagcacg tgatgaaaaa gcctgaactc 2281 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg 2341 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat 2401 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac 2461 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc 2521 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc 2581 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat 2641 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca 2701 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg 2761 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag 2821 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg 2881 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa 2941 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg 3001 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg 3061 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca 3121 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt 3181 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc 3241 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtgctacga 3301 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac 3361 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac 3421 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat 3481 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat 3541 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt 3601 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 3661 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 3721 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 3781 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 3841 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 3901 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 3961 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 4021 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 4081 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 4141 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg 4201 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 4261 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 4321 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 4381 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt 4441 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 4501 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 4561 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 4621 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 4681 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 4741 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 4801 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 4861 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 4921 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 4981 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 5041 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 5101 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 5161 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 5221 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 5281 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 5341 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 5401 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 5461 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 5521 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 5581 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 5641 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 5701 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc //