LOCUS Exported 4645 bp DNA circular SYN 08-APR-2025 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4645) TITLE Direct Submission REFERENCE 2 (bases 1 to 4645) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..4645 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind 508..524 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 621..1292 /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" CDS 1449..1475 /codon_start=1 /label=9xHis /note="9xHis affinity tag" /translation="HHHHHHHHH" CDS 1500..2216 /codon_start=1 /label=EGFP /note="enhanced GFP" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2224..2240 /codon_start=1 /label=6xHis /note="6xHis affinity tag" /translation="HHHHH" primer_bind complement(2554..2569) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(2577..2593) /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(2601..2631) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(2646..2667) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2955..3543) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3717..4574) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(join(4575..4645,1..34)) /label=AmpR promoter ORIGIN 1 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 61 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc 121 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg 181 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 241 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac 301 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca 361 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct 421 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa 481 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgccaag cttgcatgcc 541 tgcagtcaca tggtggagca cgacacactt gtctactcca aaaatatcaa agatacagtc 601 tcagaagacc aaagggcaat tgagactttt caacaaaggg taatatccgg aaacctcctc 661 ggattccatt gcccagctat ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc 721 tcctacaaat gccatcattg cgataaagga aaggccatcg ttgaagatgc ctctgccgac 781 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agacgttcca 841 accacgtctt caaagcaagt ggattgatgt gataacatgg tggagcacga cacacttgtc 901 tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga gacttttcaa 961 caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg tcactttatt 1021 gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag 1081 gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg 1141 agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat 1201 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct 1261 atataaggaa gttcatttca tttggagagg acctcgacct caacacaaca tatacaaaac 1321 aaacgaatct caagcaatca agcattctac ttctattgca gcaatttaaa tcatttcttt 1381 taaagcaaaa gcaattttct gaaaattttc accatttacg aacgatactc gagatgaccc 1441 gtggttctca tcaccatcac catcaccatc accatcacgc catggtcgac gtactagtga 1501 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 1561 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 1621 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 1681 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 1741 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 1801 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 1861 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 1921 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 1981 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 2041 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 2101 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 2161 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtcta 2221 gacatcacca tcaccatcac ggatcctgag cggccgccac cgcggtggag ctcgaatttc 2281 cccgatcgtt caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt 2341 gcgatgatta tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa 2401 tgcatgacgt tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa 2461 tacgcgatag aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca 2521 tctatgttac tagatcggga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt 2581 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt 2641 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg 2701 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg 2761 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 2821 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 2881 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 2941 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 3001 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 3061 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 3121 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg 3181 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 3241 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 3301 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 3361 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 3421 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 3481 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 3541 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 3601 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 3661 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 3721 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 3781 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 3841 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 3901 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 3961 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 4021 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 4081 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 4141 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 4201 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 4261 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 4321 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 4381 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 4441 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 4501 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 4561 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 4621 ctcatgagcg gatacatatt tgaat //