LOCUS Exported 8182 bp DNA circular SYN 03-DEC-2025 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8182) TITLE Direct Submission REFERENCE 2 (bases 1 to 8182) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..8182 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 31..47 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 57..75 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind 158..174 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature 249..3296 /label=UCOE /note="The Rps3 chromatin opening element (UCOE) is a fragment of DNA from the mouse Rps3 gene that is used in biotechnology and gene therapy to maintain stable and high-level gene expression. It functions as a ubiquitous chromatin opening element that prevents epigenetic silencing by keeping the DNA in an open, transcriptionally active state. " enhancer 3383..3686 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3687..3890 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" polyA_signal 4142..4263 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" promoter 4387..4890 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 4925..5947 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANILFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" promoter complement(6017..6035) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(6056..6072) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(6080..6096) /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(6104..6134) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(6149..6170) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6458..7046) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7220..8077) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8078..8182) /label=AmpR promoter ORIGIN 1 gcggccgcca gtggaacgaa aactcacgtt gtaaaacgac ggccagtgag cgcgcgtaat 61 acgactcact atagggcgaa ttgggtaccg ggccccccct cgaagtttat ctagggataa 121 cagggtaatg ataaacattt cggataacaa tttcacacag gaaacagcta tgaccatgat 181 tacgccaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt 241 cgccctttac cactaagcca tctctccagc cctgagtcat ggttttagtg tgagaggcat 301 cattgaattt tctgagcacg gccatcaggg tagctggcac aggtcttcag atacaaggag 361 atagttataa gaaggcagcc atggctgtgg tgcactagaa atggagaaac agcttcatca 421 ggtgacagac cagtctgact ctgtcccatg attagaagcc atcttgttac aaggtcaaaa 481 taagttcatt cctgttttct gtaacacttg ggtttgatcc tgtcgtcaac ccattttctg 541 gaatttgaca tgttccatac tccattatac cctgacttcc accctgataa gatgttctgc 601 caagttcctg tgtagccaac attcccctgg aaatctctct tcccttggaa accacctagt 661 cttagaaatt ttgagttata taaattccac ttctatgttt gatgctattc tttaaaactc 721 cactttaggg agatagccct gtctgataga aaataaaact tgcttaattt gtctaaaaga 781 ttttaagtaa tagtttttac ttttgttccg tgggattagt acagggtgaa acagactccc 841 gtgtttccag tgtgaagtga gccacacact gcagtacaag ttatatcagc aggttctgcc 901 tctgcgcaat gaacttttgc ttgtgtggac atcagggtct gtgtgaaggg aaggtcctat 961 ggcctagttt tatactattc aacagtctgt ccccgaagcc ctggtgcttt attattttga 1021 caagcccctg ctgctggtat tccaccctgc tgcgagtcaa aaaagttcct gtctcggaaa 1081 aacaaaacaa aacaaaacaa ccaaaaaata aatttttttt tcccacaggt tctagtggag 1141 gtgctcacta ccagaaatcc tacaaataag cccatctcat ggatcagggt ttacctttgt 1201 aataatatta aatctgtgtg catgtgcgca cgcatgtgtt ttatgcttgc atatatgtat 1261 acgcagccat ggttttctac tgtcccactc actctgtaac ttactgagcc atccagctgg 1321 tcctctaaat acatttcaat gaaagttttc attagcgtga acgtgaaggt ggtaaaatct 1381 gttagtgtgt gcttatgcct gtggtttgca cctctagtct gaaggttgct cttttcaaat 1441 tttttattta tttacgtttt tacttctgag tcagaaactc ataaaggcca tggcctcgaa 1501 ttcgctatgt agtcaacgat gaccttaaac ttgtgaccct ctacttcgtt agtgctggaa 1561 ccccaagctt gctgagtaca gagcactttc agaccggaac tagatgtcta cttcctgttc 1621 cgcctacatt acaggttgct aggttacacc ccccctacgc cgttttagac gcaaaacttc 1681 atttcccatg caaaacttca tttcccatga acacttgcaa gggtcgccgc gctgcgcggc 1741 gtcattgctc ccgccctata tacctacttc cgcccgcgag ccacttcctt tcctttcagc 1801 ggcgcgcggc tgcaagatgg cggtgcagat ttccaagaag aggaaggtaa gcgtctgggc 1861 ccggttcggg agtccgccgc gggttctaca agtgccaggg aggcctgtgg ctccgtaatc 1921 agtcctgtgg agcgtctggg gccgcctgcc gtctcttcga gcctcggatg gccgtagatt 1981 gtgtattggg ccggagccgg gcgagtgctg tgtgcctggg caagggaggg acaaactcct 2041 cgagttctgg accgactcga acaccgggcg cctccagttc cggactagac acctttgagc 2101 gtttcttggt ctccataata gtaatcctgt ggcacagtta gagggcgtgt gccatcagat 2161 ctagtccagt ctctttagta agtgaagttt agcagtccct tctcttagtc gcgtgatcct 2221 gcaagtggcc atagttgaaa gcctacttac tgactgctgc cgtgttcact cgggacccgg 2281 agctgcagcg tccctgtggt tatcatttca tgggggaaaa gtgtgcaggt tgccaggttt 2341 agaaatagat ggtctgtcgt ttgtgcttat gcacacagat gataaacctg ttttgagtca 2401 ggattcctct cctatccgag gtacaactta cagtcccagc tgtacatgtg ctacttggag 2461 acagattttt ctttgtctct tgggtgtaga ttatgccgta gagcccttcg atgaagaggt 2521 gatgacgagt ctgagtagga agtgttgtct ttgtccaaga tgcctcacta tgctgcgttc 2581 tgtggcacag ctgaaagcac tgtggtcaaa agaaacttcc taaagatgac caagaggcat 2641 ttgtctgaga agggttgctg cttttctgta gggccattgg gcttgctctg actaaccctg 2701 tcttcacctc agaggtaact tgtttccttt ggttcagttt gtagctgatg gcatcttcaa 2761 agctgagctg aatgaatttc tcactcggga gctggctgaa gatggctact ctggagttga 2821 agtccgagtt acaccaacca ggacagaaat cattatttta gccaccaggt agaaatacca 2881 ttgattgtca cctgtaaata ttgtgtgtac tgagatgctg tgtaaacttg ggccaaccaa 2941 gcagtaaatc tggcctcagt gggtgtaact gctttgttag aactgcattt gggaagaact 3001 taccttccat ttaacctgtg tgctggcgtt gtggtgggcg gcaggtggga tcttgagtaa 3061 atggttgcgc ttcccctcta caggacacag aatgttcttg gggagaaggg tcgtcggatc 3121 agagagttga ccgcagttgt ccagaagcgc tttggcttcc ctgaaggcag cgtagaggtg 3181 agttcctctg ctttatctcc cgggggtttt agactgagtt gggatgtggc ttctgctata 3241 gaattgtact tctgaaaacc tgacatggcc agtgacagtc acaggtactt gatgctaagg 3301 gcgaattctg cagatgggcc gtacgaattt aatagtaatc aattacgggg tcattagttc 3361 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac 3421 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa 3481 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag 3541 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc 3601 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct 3661 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg 3721 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt 3781 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga 3841 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga 3901 accgtcagat ccgctagaat tcgagctccc tgcaggttag ttaagttaac ggccggccgc 3961 tagcgcgatc gcgagctcgg tacccgggga tcctctagag tcgacctgca ggcatgcaag 4021 ctggccgcga ctctagatca taatcagcca taccacattt gtagaggttt tacttgcttt 4081 aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt 4141 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac 4201 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc 4261 ttaaagctat ccatcacact ggcggccgaa atctagaagc ttatcgatat ggcgcgccaa 4321 ttgttaggcc gcattcgcca atcaagtaag ctaagatctg gcccaatggg ccgtacgaat 4381 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc 4441 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg 4501 gccggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct 4561 cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg 4621 gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg 4681 gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg 4741 ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc 4801 cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg 4861 ttctcctctt cctcatctcc gggcctttcg acggtaccgc gggcccgggg gatccactag 4921 tactatgaaa aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt 4981 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt 5041 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa 5101 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga 5161 cattggggag ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac 5221 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctc cagccggtcg cggaggccat 5281 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca 5341 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt 5401 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga 5461 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc atgcggattt 5521 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga 5581 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ctcttctgga ggccgtggtt 5641 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc 5701 gccgcgcctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt 5761 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc 5821 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga 5881 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc 5941 aaaggaatag agtagatgcc gaccgggaac cagttaacgt cgacctcgag ggggggcccg 6001 gtaccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat 6061 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 6121 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 6181 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 6241 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 6301 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 6361 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 6421 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 6481 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 6541 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 6601 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 6661 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 6721 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 6781 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 6841 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 6901 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 6961 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 7021 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 7081 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 7141 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 7201 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 7261 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 7321 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 7381 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 7441 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 7501 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 7561 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 7621 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 7681 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 7741 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 7801 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 7861 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 7921 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 7981 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 8041 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 8101 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 8161 ataaacaaat aggggttccg cg //