LOCUS Exported 3508 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pMLM36 (aka pBR-UV5-GP-FD2) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3508) TITLE OPEN plasmids REFERENCE 2 (bases 1 to 3508) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3508 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 112..303 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" primer_bind complement(297..319) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 405..545 /label=bom /note="basis of mobility region from pBR322" primer_bind 460..479 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(560..577) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(731..1319) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(811..830) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1490..2350) /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 2113..2132 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2351..2455) /gene="bla" /label=AmpR promoter rep_origin complement(2481..2936) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2618..2639) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2830..2849 /label=F1ori-R /note="F1 origin, reverse primer" CDS complement(3114..3137) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(3428..3450) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3439..3455 /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(3463..3493) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" ORIGIN 1 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc 61 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa 121 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg 181 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac 241 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc 301 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga 361 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag 421 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac 481 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca 541 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 601 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 661 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 721 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 781 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 841 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 901 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 961 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 1021 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 1081 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 1141 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga 1201 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 1261 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 1321 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 1381 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 1441 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 1501 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 1561 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 1621 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 1681 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 1741 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 1801 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 1861 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 1921 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 1981 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 2041 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 2101 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 2161 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 2221 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 2281 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 2341 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 2401 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 2461 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa 2521 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa 2581 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact 2641 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc 2701 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa 2761 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc 2821 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt 2881 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca 2941 ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt 3001 acgccaggat cttcagcgga gcgggacttg tcggcctttt tctagtctct agttactact 3061 gtgcatgtct tcttacttag gatcctcact tagaagacct ccccgggtct agatttgtca 3121 tcatcgtctt tgtagtcagc ggccgccaaa gcttggattt ttctcagggc cttttgcaca 3181 atgtatttcc taataaaact ttgcttgaca ggttccgtca tcgtactacc caattgtgtt 3241 ttttcaaaat tcttgcaggt ttcgatggct tcttgtaata agctgcgttg ctcctctgga 3301 gaaaagattt ggttaatttg gttgaggacg tttggtacgg tattttgtga tgattgctgt 3361 tgcaaagggt tgttgtccgc acttgaattg ggaggttgca tttgctgctg ttgaggcata 3421 gctgtttcct gtgtgaaatt gttatccgct cacaagtcga cacattatac gagccggaag 3481 cataaagtgt aaagcccgga attcccgt //