LOCUS       sequence-149930-        6163 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 6163)
  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 6163)
  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
FEATURES             Location/Qualifiers
     source          1..6163
                     /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"
     rep_origin      1389..1844
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1476..1495)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1955..2543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2444..2463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2697..2714
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2880..2902
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3041..3060
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(3240..4118)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4145..4429
                     /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     4352..4371
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             4464..5249
                     /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 tcccttcctt
     1501 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1561 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     1621 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1681 taatagtgga ctcttgttcc aaacttgaac aacactcaac cctatctcgg gctattcttt
     1741 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     1801 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttaaaagg atctaggtga
     1861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2041 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2101 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2581 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2641 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2701 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     2761 gcggtatttc acaccgcata gggtcatggc tgcgccccga cacccgccaa cacccgctga
     2821 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     2881 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcaagg
     2941 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     3001 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     3061 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     3121 tctgctcatg tttgacagct tatcatcgat gcataatgtg cctgtcaaat ggacgaagca
     3181 gggattctgc aaaccctatg ctactccgtc aagccgtcaa ttgtctgatt cgttaccaat
     3241 tatgacaact tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc
     3301 gggctggccc cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca
     3361 acattgcgac cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg
     3421 ctgatacgtt ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga
     3481 tgtgacagac gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg
     3541 ctgtctgcca ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt
     3601 ggatggagcg actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt
     3661 atcgccagca gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac
     3721 aggtcgctga aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat
     3781 attgacggcc agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg
     3841 tgataccatt cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac
     3901 agcaaaatat cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga
     3961 ccgcgaatgg tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa
     4021 tcgagataac cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag
     4081 tatcccggca gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc
     4141 agagaagaaa ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg
     4201 ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg
     4261 accaaagcca tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac
     4321 attgattatt tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta
     4381 gcggatccta cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt
     4441 tgggctagca ggaggaattc accatgggta ctgcactcgc aacgctggct gggaagctgg
     4501 ctgaacgtgt cggcatggat tctgtcgacc cacaggaact gatcaccact cttcgccaga
     4561 cggcatttaa aggtgatgcc agcgatgcgc agttcatcgc attactgatc gttgccaacc
     4621 agtacggcct taatccgtgg acgaaagaaa tttacgcctt tcctgataag cagaatggca
     4681 tcgttccggt ggtgggcgtt gatggctggt cccgcatcat caatgaaaac cagcagtttg
     4741 atggcatgga ctttgagcag gacaatgaat cctgtacatg ccggatttac cgcaaggacc
     4801 gtaatcatcc gatctgcgtt accgaatgga tggatgaatg ccgccgcgaa ccattcaaaa
     4861 ctcgcgaagg cagagaaatc acggggccgt ggcagtcgca tcccaaacgg atgttacgtc
     4921 ataaagccat gattcagtgt gcccgtctgg ccttcggatt tgctggtatc tatgacaagg
     4981 atgaagccga gcgcattgtc gaaaatactg catacactgc agaacgtcag ccggaacgcg
     5041 acatcactcc ggttaacgat gaaaccatgc aggagattaa cactctgctg atcgccctgg
     5101 ataaaacatg ggatgacgac ttattgccgc tctgttccca gatatttcgc cgcgacattc
     5161 gtgcatcgtc agaactgaca caggccgaag cagtaaaagc tcttggattc ctgaaacaga
     5221 aagccgcaga gcagaaggtg gcagcatgaa gtcggtgtgg agtgaaacga tgaagaaaaa
     5281 tcgcgctttt ttgaagtggg cagggggcaa gtatcccctg cttgatgata ttaaacggca
     5341 tttgcccaag ggcgaatgtc tggttgagcc ttttgtaggt gccgggtcgg tgtttctcaa
     5401 caccgacttt tctcgttata tccttgccga tatcaatagc gacctgatca gtctctataa
     5461 cattgtgaag atgcgtactg atgagtacgt acaggccgca cgcgagctgt ttgttcccga
     5521 aacaaattgc gccgaggttt actatcagtt ccgcgaagag ttcaacaaaa gccaggatcc
     5581 gttccgtcgg gcggtactgt ttttatattt gaaccgctac ggttacaacg gcctgtgtcg
     5641 ttacaatctg cgcggtgagt ttaacgtgcc gttcggccgc tacaaaaaac cctatttccc
     5701 ggaagcagag ttgtatcact tcgctgaaaa agcgcagaat gcctttttct attgtgagtc
     5761 ttacgccgat agcatggcgc gcgcagatga tgcatccgtc gtctattgcg atccgcctta
     5821 tgcaccgctg tctgcgaccg ccaactttac ggcgtatcac acaaacagtt ttacgcttga
     5881 acaacaagcg catctggcgg agatcgccga aggtctggtt gagcgccata ttccagtgct
     5941 gatctccaat cacgatacga tgttaacgcg tgagtggtat cagcgcgcaa aattgcatgt
     6001 cgtcaaagtt cgacgcagta taagcagcaa cggcggcaca cgtaaaaagg tggacgaact
     6061 gctggctttg tacaaaccag gagtcgtttc acccgcgaaa aaataataag cttggctgtt
     6121 ttggcggatg agagaagatt ttcagcctga tacagattaa atc
//
