LOCUS       sequence-149931-        7337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  TM-MAGE strain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7337)
  AUTHORS   Lennen RM, Nilsson Wallin AI, Pedersen M, Bonde M, Luo H, Herrgard 
            MJ, Sommer MO
  TITLE     Transient overexpression of DNA adenine methylase enables efficient 
            and mobile genome engineering with reduced off-target effects.
  JOURNAL   Nucleic Acids Res. 2016 Feb 29;44(4):e36. doi: 10.1093/nar/gkv1090. 
            Epub 2015 Oct 22.
  PUBMED    26496947
REFERENCE   2  (bases 1 to 7337)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2016-02-29"; volume: "44(4):e36. doi"; pages: " 
            10.1093/nar/gkv1090. Epub 2015 Oct 22"
COMMENT     Sequence Label: pMA7-SacB
FEATURES             Location/Qualifiers
     source          1..7337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      171..257
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      349..376
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        395..486
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(487..555,556..1347)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(705..724)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             join(1659..1745,1746..3080)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     rep_origin      3129..3717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3618..3637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3871..3888
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4054..4076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4215..4234
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4414..5292)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5319..5603
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     5526..5545
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             5638..6423
                     /codon_start=1
                     /gene="bet"
                     /product="single-stranded DNA binding recombinase in the 
                     lambda Red system"
                     /label=Beta
                     /note="mediates pairing of a single-stranded DNA with a 
                     complementary target"
                     /translation="MGTALATLAGKLAERVGMDSVDPQELITTLRQTAFKGDASDAQFI
                     ALLIVANQYGLNPWTKEIYAFPDKQNGIVPVVGVDGWSRIINENQQFDGMDFEQDNESC
                     TCRIYRKDRNHPICVTEWMDECRREPFKTREGREITGPWQSHPKRMLRHKAMIQCARLA
                     FGFAGIYDKDEAERIVENTAYTAERQPERDITPVNDETMQEINTLLIALDKTWDDDLLP
                     LCSQIFRRDIRASSELTQAEAVKALGFLKQKAAEQKVAA"
ORIGIN
        1 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
       61 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
      121 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
      181 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
      241 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
      301 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
      361 tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca
      421 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      481 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      541 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
      601 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
      661 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
      721 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
      781 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
      841 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
      901 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
      961 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1021 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1081 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1141 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1201 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1261 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1321 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1381 attgatttac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1441 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcctagggat
     1501 aacagggtaa tgtgtaaagg gcaaagtgta tactttggcg tcacccctta catattttag
     1561 gtcttttttt attgtgcgta actaacttgc catcttcaaa caggagggct ggaagaagca
     1621 gaccgctaac acagtacata aaaaaggaga catgaacgat gaacatcaaa aagtttgcaa
     1681 aacaagcaac agtattaacc tttactaccg cactgctggc aggaggcgca actcaagcgt
     1741 ttgcgaaaga aacgaaccaa aagccatata aggaaacata cggcatttcc catattacac
     1801 gccatgatat gctgcaaatc cctgaacagc aaaaaaatga aaaatatcaa gttcctgaat
     1861 tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg cctggacgtt tgggacagct
     1921 ggccattaca aaacgctgac ggcactgtcg caaactatca cggctaccac atcgtctttg
     1981 cattagccgg agatcctaaa aatgcggatg acacatcgat ttacatgttc tatcaaaaag
     2041 tcggcgaaac ttctattgac agctggaaaa acgctggccg cgtctttaaa gacagcgaca
     2101 aattcgatgc aaatgattct atcctaaaag accaaacaca agaatggtca ggttcagcca
     2161 catttacatc tgacggaaaa atccgtttat tctacactga tttctccggt aaacattacg
     2221 gcaaacaaac actgacaact gcacaagtta acgtatcagc atcagacagc tctttgaaca
     2281 tcaacggtgt agaggattat aaatcaatct ttgacggtga cggaaaaacg tatcaaaatg
     2341 tacagcagtt catcgatgaa ggcaactaca gctcaggcga caaccatacg ctgagagatc
     2401 ctcactacgt agaagataaa ggccacaaat acttagtatt tgaagcaaac actggaactg
     2461 aagatggcta ccaaggcgaa gaatctttat ttaacaaagc atactatggc aaaagcacat
     2521 cattcttccg tcaagaaagt caaaaacttc tgcaaagcga taaaaaacgc acggctgagt
     2581 tagcaaacgg cgctctcggt atgattgagc taaacgatga ttacacactg aaaaaagtga
     2641 tgaaaccgct gattgcatct aacacagtaa cagatgaaat tgaacgcgcg aacgtcttta
     2701 aaatgaacgg caaatggtac ctgttcactg actcccgcgg atcaaaaatg acgattgacg
     2761 gcattacgtc taacgatatt tacatgcttg gttatgtttc taattcttta actggcccat
     2821 acaagccgct gaacaaaact ggccttgtgt taaaaatgga tcttgatcct aacgatgtaa
     2881 cctttactta ctcacacttc gctgtacctc aagcgaaagg aaacaatgtc gtgattacaa
     2941 gctatatgac aaacagagga ttctacgcag acaaacaatc aacgtttgcg ccaagcttcc
     3001 tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga cagcatcctt gaacaaggac
     3061 aattaacagt taacaaataa ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3121 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3181 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3241 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3301 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3361 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3421 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3481 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3541 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3601 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3661 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3721 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     3781 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     3841 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     3901 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagggtca
     3961 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4021 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4081 caccgtcatc accgaaacgc gcgaggcagc aaggagatgg cgcccaacag tcccccggcc
     4141 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     4201 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     4261 gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac agcttatcat
     4321 cgatgcataa tgtgcctgtc aaatggacga agcagggatt ctgcaaaccc tatgctactc
     4381 cgtcaagccg tcaattgtct gattcgttac caattatgac aacttgacgg ctacatcatt
     4441 cactttttct tcacaaccgg cacggaactc gctcgggctg gccccggtgc attttttaaa
     4501 tacccgcgag aaatagagtt gatcgtcaaa accaacattg cgaccgacgg tggcgatagg
     4561 catccgggtg gtgctcaaaa gcagcttcgc ctggctgata cgttggtcct cgcgccagct
     4621 taagacgcta atccctaact gctggcggaa aagatgtgac agacgcgacg gcgacaagca
     4681 aacatgctgt gcgacgctgg cgatatcaaa attgctgtct gccaggtgat cgctgatgta
     4741 ctgacaagcc tcgcgtaccc gattatccat cggtggatgg agcgactcgt taatcgcttc
     4801 catgcgccgc agtaacaatt gctcaagcag atttatcgcc agcagctccg aatagcgccc
     4861 ttccccttgc ccggcgttaa tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc
     4921 ttcatccggg cgaaagaacc ccgtattggc aaatattgac ggccagttaa gccattcatg
     4981 ccagtaggcg cgcggacgaa agtaaaccca ctggtgatac cattcgcgag cctccggatg
     5041 acgaccgtag tgatgaatct ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac
     5101 aaattctcgt ccctgatttt tcaccacccc ctgaccgcga atggtgagat tgagaatata
     5161 acctttcatt cccagcggtc ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg
     5221 cgttaaaccc gccaccagat gggcattaaa cgagtatccc ggcagcaggg gatcattttg
     5281 cgcttcagcc atacttttca tactcccgcc attcagagaa gaaaccaatt gtccatattg
     5341 catcagacat tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac
     5401 cccgcttatt aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa
     5461 caaaagtgtc tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact
     5521 ttgctatgcc atagcatttt tatccataag attagcggat cctacctgac gctttttatc
     5581 gcaactctct actgtttctc catacccgtt tttttgggct agcaggagga attcaccatg
     5641 ggtactgcac tcgcaacgct ggctgggaag ctggctgaac gtgtcggcat ggattctgtc
     5701 gacccacagg aactgatcac cactcttcgc cagacggcat ttaaaggtga tgccagcgat
     5761 gcgcagttca tcgcattact gatcgttgcc aaccagtacg gccttaatcc gtggacgaaa
     5821 gaaatttacg cctttcctga taagcagaat ggcatcgttc cggtggtggg cgttgatggc
     5881 tggtcccgca tcatcaatga aaaccagcag tttgatggca tggactttga gcaggacaat
     5941 gaatcctgta catgccggat ttaccgcaag gaccgtaatc atccgatctg cgttaccgaa
     6001 tggatggatg aatgccgccg cgaaccattc aaaactcgcg aaggcagaga aatcacgggg
     6061 ccgtggcagt cgcatcccaa acggatgtta cgtcataaag ccatgattca gtgtgcccgt
     6121 ctggccttcg gatttgctgg tatctatgac aaggatgaag ccgagcgcat tgtcgaaaat
     6181 actgcataca ctgcagaacg tcagccggaa cgcgacatca ctccggttaa cgatgaaacc
     6241 atgcaggaga ttaacactct gctgatcgcc ctggataaaa catgggatga cgacttattg
     6301 ccgctctgtt cccagatatt tcgccgcgac attcgtgcat cgtcagaact gacacaggcc
     6361 gaagcagtaa aagctcttgg attcctgaaa cagaaagccg cagagcagaa ggtggcagca
     6421 tgaagtcggt gtggagtgaa acgatgaaga aaaatcgcgc ttttttgaag tgggcagggg
     6481 gcaagtatcc cctgcttgat gatattaaac ggcatttgcc caagggcgaa tgtctggttg
     6541 agccttttgt aggtgccggg tcggtgtttc tcaacaccga cttttctcgt tatatccttg
     6601 ccgatatcaa tagcgacctg atcagtctct ataacattgt gaagatgcgt actgatgagt
     6661 acgtacaggc cgcacgcgag ctgtttgttc ccgaaacaaa ttgcgccgag gtttactatc
     6721 agttccgcga agagttcaac aaaagccagg atccgttccg tcgggcggta ctgtttttat
     6781 atttgaaccg ctacggttac aacggcctgt gtcgttacaa tctgcgcggt gagtttaacg
     6841 tgccgttcgg ccgctacaaa aaaccctatt tcccggaagc agagttgtat cacttcgctg
     6901 aaaaagcgca gaatgccttt ttctattgtg agtcttacgc cgatagcatg gcgcgcgcag
     6961 atgatgcatc cgtcgtctat tgcgatccgc cttatgcacc gctgtctgcg accgccaact
     7021 ttacggcgta tcacacaaac agttttacgc ttgaacaaca agcgcatctg gcggagatcg
     7081 ccgaaggtct ggttgagcgc catattccag tgctgatctc caatcacgat acgatgttaa
     7141 cgcgtgagtg gtatcagcgc gcaaaattgc atgtcgtcaa agttcgacgc agtataagca
     7201 gcaacggcgg cacacgtaaa aaggtggacg aactgctggc tttgtacaaa ccaggagtcg
     7261 tttcacccgc gaaaaaataa taagcttggc tgttttggcg gatgagagaa gattttcagc
     7321 ctgatacaga ttaaatc
//
