LOCUS       sequence-176849-       11390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pMAD2 derivative designed for chromosomal gene ectopic 
            complementation in Staphylococcus aureus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11390)
  TITLE     pMAD derivatives
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11390)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pECTO
FEATURES             Location/Qualifiers
     source          1..11390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      7101..7172
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             8947..9138
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(9132..9154)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    9240..9380
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     9295..9314
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(9395..9412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(9566..10154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(9646..9665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10325..11116,11117..11185))
                     /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     10948..10967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(11186..11290)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     11358..11376
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gaattaattc tcatgtttga cagcttatca tcgattcaca aaaaataggc acacgaaaaa
       61 caagttaagg gatgcagttt atgcatccct taacttactt attaaataat ttatagctat
      121 tgaaaagaga taagaattgt tcaaagctaa tattgtttaa atcgtcaatt cctgcatgtt
      181 ttaaggaatt gttaaattga ttttttgtaa atattttctt gtattctttg ttaacccatt
      241 tcataacgaa ataattatac ttttgtttat ctttgtgtga tattcttgat ttttttctac
      301 ttaatctgat aagtgagcta ttcactttag gtttaggatg aaaatattct cttggaacca
      361 tacttaatat agaaatatca acttctgcca ttaaaagtaa tgccaatgag cgttttgtat
      421 ttaataatct tttagcaaac ccgtattcca cgattaaata aatctcatta gctatactat
      481 caaaaacaat tttgcgtatt atatccgtac ttatgttata aggtatatta ccatatattt
      541 tataggattg gtttttagga aatttaaact gcaatatatc cttgtttaaa acttggaaat
      601 tatcgtgatc aacaagttta ttttctgtag ttttgcataa tttatggtct atttcaatgg
      661 cagttacgaa attacacctc tttactaatt caagggtaaa atggcctttt cctgagccga
      721 tttcaaagat attatcatgt tcatttaatc ttatatttgt cattatttta tctatattat
      781 gttttgaagt aataaagttt tgactgtgtt ttatattttt ctcgttcatt ataaccctct
      841 ttaatttggt tatatgaatt ttgcttatta acgattcatt ataaccactt attttttgtt
      901 tggttgataa tgaactgtgc tgattacaaa aatactaaaa atgcccatat tttttcctcc
      961 ttataaaatt agtataatta tagcacgagc tctgataaat atgaacatga tgagtgatcg
     1021 ttaaatttat actgcaatcg gatgcgatta ttgaataaaa gatatgagag atttatctaa
     1081 tttctttttt cttgtaaaaa aagaaagttc ttaaaggttt tatagttttg gtcgtagagc
     1141 acacggttta acgacttaat tacgaagtaa ataagtctag tgtgttagac tttatgaaat
     1201 ctatatacgt ttatatatat ttattatccg gaggtgtagc atgtctcatt caattttgag
     1261 ggttgccaga gttaaaggat caagtaatac aaacgggata caaagacata atcaaagaga
     1321 gaataaaaac tataataata aagacataaa tcatgaggaa acatataaaa attatgattt
     1381 gattaacgca caaaatataa agtataaaga taaaattgat gaaacgattg atgagaatta
     1441 ttcagggaaa cgtaaaattc ggtcagatgc aattcgacat gtggacggac tggttacaag
     1501 tgataaagat ttctttgatg atttaagcgg agaagaaata gaacgatttt ttaaagatag
     1561 cttggagttt ctagaaaatg aatacggtaa ggaaaatatg ctgtatgcga ctgtccatct
     1621 ggatgaaaga gtcccacata tgcactttgg ttttgtccct ttaacagagg acgggagatt
     1681 gtctgcaaaa gaacagttag gcaacaagaa agactttact caattacaag atagatttaa
     1741 tgagtatgtg aatgagaaag gttatgaact tgaaagaggc acgtccaaag aggttacaga
     1801 acgagaacat aaagcgatgg atcagtacaa gaaagatact gtatttcata aacaggaact
     1861 gcaagaagtt aaggatgagt tacagaaggc aaataagcag ttacagagtg gaatagagca
     1921 tatgaggtct acgaaaccct ttgattatga aaatgagcgt acaggtttgt tctctggacg
     1981 tgaagagact ggtagaaaga tattaactgc tgatgaattt gaacgcctgc aagaaacaat
     2041 ctcttctgca gaacggattg ttgatgatta cgaaaatatt aagagcacag actattacac
     2101 agaaaatcaa gaattaaaaa aacgtagaga gagtttgaaa gaagtagtga atacatggaa
     2161 agaggggtat cacgaaaaaa gtaaagaggt taataaatta aagcgagaga atgatagttt
     2221 gaatgagcag ttgaatgtat cagagaaatt tcaagatagt acagtgactt tatatcgtgc
     2281 tgcgagggcg aatttccctg ggtttgagaa agggtttaat aggcttaaag agaaattctt
     2341 taatgattcc aaattcgagc gtgtgggaca gtttatggat gttgtacagg ataatgtcca
     2401 gaaggtcgat agaaagcgtg agaaacagcg tacagacgat ttagagatgt agaggtactt
     2461 ttatgccgag aaaacttttt gcgtgtgaca gtccttaaaa tatacttaga gcgtaagcga
     2521 aagtagtagc gacagctatt aactttcggt tgcaaagctc taggattttt aatggacgca
     2581 gcgcatcaca cgcaaaaagg aaattggaat aaatgcgaaa tttgagatgt taattaaaga
     2641 cctttttgag gtcttttttt cttagatttt tggggttatt taggggagaa aacatagggg
     2701 ggtactacga cctcccccct aggtgtccat tgtccattgt ccaaacaaat aaataaatat
     2761 tgggttttta atgttaaaag gttgtttttt atgttaaagt gaaaaaaaca gatgttggga
     2821 ggtacagtga tggttgtaga tagaaaagaa gagaaaaaag ttgctgttac tttaagactt
     2881 acaacagaag aaaatgagat attaaataga atcaaagaaa aatataatat tagcaaatca
     2941 gatgcaaccg gtattctaat aaaaaaatat gcaaaggagg aatacggtgc attttaaaca
     3001 aaaaaagata gacagcactg gcatgctgcc tatctatgac taaattttgt taaatgtatt
     3061 agcaccgtta ttatatcatg agcgaaaatg taataaaaga aactgaaaac aagaaaaatt
     3121 caagaggacg taattggaca tttgttttat atccagaatc agcaaaagcc gagtggttag
     3181 agtatttaaa agagttacac attcaatttg tagtgtctcc attacatgat agggatactg
     3241 atacagaaga taggatgaaa aaagagcatt atcatattct agtgatgtat gagggtaata
     3301 aatcttatga acagataaaa ataattacag aagaattgaa tgcgactatt ccgcagattg
     3361 caggaagtgt gaaaggtctt gtgagatata tgcttcacat ggacgatcct aataaattta
     3421 aatatcaaaa agaagatatg atagtttatg gcggtgtaga tgttgatgaa ttattaaaga
     3481 aaacaacaac agatagatat aaattaatta aagaaatgat tgagtttatt gatgaacaag
     3541 gaatcgtaga atttaagagt ttaatggatt atgcaatgaa gtttaaattt gatgattggt
     3601 tcccgctttt atgtgataac tcggcgtatg ttattcaaga atatataaaa tcaaatcggt
     3661 ataaatctga ccgatagatt ttgaatttaa gagtgtcaca agacactctt ttttcgcacc
     3721 aacgaaaact ggtttaagcc gactgcgcaa aagacataat cgataagctt taatgcggta
     3781 gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct aacaatgcgc
     3841 tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg gttatgccgg
     3901 tactgccggg cctcttgcgg gatcgaggcc tttagcatat tatgttgcca actgtcggaa
     3961 cgagacttct ctaaaccttc ccggcttcat catgctctct taacactgct acatcaaccc
     4021 cttgaatcct caattcacct cctcgaaaac atttcccatc aatcatattc tggtatatct
     4081 tatcttctgg cagtgacagc gtcacttcgt aatcattatg attgatgata atcaaatact
     4141 tccattcatc agtctctctt tgttgcacct ctacattttc agctacttca agaatgggat
     4201 taatatgatg tttagcgaaa acctgttcta aaagcctgcc taaataatta ctatctggat
     4261 aagtaccgac gtaaatcccc tctcctttac cgtagcagtt acgtgtaacc gccggaagtc
     4321 ctgcatacca atccccctta aatgtcgcta gaggttctgc cccttctaat cggattatgt
     4381 ccgcccacgt cgtacaatca tattccccat cgttactata tattttgttt acctttgttt
     4441 ctgggtatgg tacaaattcc tcaacaaaaa tccccaaaat atctcgcaga ggaccaggat
     4501 atccgcctag atgtacacgg tcattttcat ctacaatgcc actgaagaaa ctgacaatca
     4561 aagtgccacc gttagcaaca aattgccgta agttttcatc ttctccctct ttaaccatat
     4621 ataacattgg agcaataact actttgtatt ttgttagatc atcagatggc cttacaaaat
     4681 cgacagcaat attacgttta tataattccc tataataagc ttcaactata ggaatatatc
     4741 ttagtttatt atgtggtttg gaacttagtt cgacagccca ccagttttcc caatcaaaaa
     4801 tgatcgcgac ctctgccttg attctagatc cgaccaaaca atctaacttt ttcagctctt
     4861 gccctaactg tgtaacttcc ctataaattc tattattctc gttcaaaaag tggggcacca
     4921 ttgcaccgtg gaatttttca gctcctgctc tactttgacg ccactggaaa aacataatac
     4981 catctgcacc acgggcaata gttgcataac tccatagacg cattacacct ggcggttttg
     5041 gaacattaat atcgcgccag ttaacatgtg aggttacctg ctccatcaaa ataaacggtt
     5101 gaccttttct taaactacgc ataaggtcat tcatcatggc gtgctgaatt ggcaagccct
     5161 ctctgggatc aggatatgag tcccatgtca caatatctac atgctgagcc cattgaaaat
     5221 agtttaacgg tttgaatgaa cccatgaaat tagttgatac tggaatatct ggtgttacct
     5281 cacgtaaaat ttccttttct gttaaaaaca acttgagaat tgagtcattc ataaaacggt
     5341 agtaatcaag ttcttgggat ggattaataa aagttggtgc ctttctaggg ggattaattt
     5401 catcccaatg attgtatcgc tgtccccaaa agtttgtacc ccaacgttca tttaattcat
     5461 cgattgtttt atatctttcc tttagccact ttctaaacgc gacagcacaa ttctcacaaa
     5521 aacacttgga aacgtgacat gcatactcat tattaacatg ccacattttg agtgccggat
     5581 gatttttata ccgttctgct atagccctca caagtctctt tatgtgcgta attaattgag
     5641 gatgattagg acaataatgt tgtctactgc caaacgagag aatgacaccg ctttcatcga
     5701 tcggcaaaga atctggatac ttttttacaa accaagctgg agtagttgca gtcgccgtcc
     5761 ccaagttaat ataaacaccg tggtcatata gtatatctat aaccttgtct agccattcaa
     5821 agtcgaacac tccatcagac ggttcgatct tgctccaact gaaaatccct aaagatacta
     5881 aattcacccc cgctttttgc atcaacttag catcttcata ccaaatttcc tctggccatt
     5941 gctctgggtt ataatctcct ccgtaacaaa ttgaggataa cacattcata attattcccc
     6001 ctagctaatt ttcgtttaat tataaattaa gttaaaattt aggtacctac gtaggatcga
     6061 tccgacctta attatattat agtcccaata taatcattta tcaactcttt tacacttaaa
     6121 tttcctaata tttgtaaata tttttttgaa taaaaaaatc tagctaatgt tacgttacac
     6181 attaactaga cagatctatc gatgcatgcc atggtacccg ggagctcgaa ttcgagaccg
     6241 ctagcgcagt ttgatcaatt aggccgtgaa aactattata tgtataaaaa gtttggtgtt
     6301 ttaccagaaa tggaatacga aatggaagaa gttaaagaaa tcgagcaata tattaaagag
     6361 caagatgaag ctgaactata ggggcgataa catgactttc gaagaaaagc taagtcaaat
     6421 gtacaatgaa attgcaaatg aaatcagtgg aatgatacca gtagaatggg aaaaagtata
     6481 tacaattgcc tacgtagatg atgaaggtgg agaggttgtt tttaattata ctaaaccagg
     6541 aagtgaagat ttgaattatt attcagatat tcctaaagat tgcaatgtct caaaagatat
     6601 ttttaagaat tcatggttta aagtttatcg aatgtttgat gagttaagag aaacttttaa
     6661 aaaagaagat ttagaaccgt ggacatcatg tgaatttgac tttacaagaa agggaaattt
     6721 aaaagtatca tttgattata tagattggat taaattaggt tttggcccat caggaaagga
     6781 aaactactat atgtacaaaa aatttggtat tttaccagat atggaatatg aaatggaaga
     6841 aattcgagca gtagagaagt atgttaaaga gcaagagtag cagacatgtt ataaaagact
     6901 gtgcaaaatc accctcgttt tacatttgat tcaaagaaga aggtaaaaga taagattatt
     6961 tgcaacttaa aaagtcaatt agcttatcgg tatccataca tcatggataa atgagttcaa
     7021 ctaattaaca aatcacgata taaatttatc gattgataag tcgacgtctc cccatgcgag
     7081 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcggaagac tgggcctttc
     7141 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gattgtcatt aattgaaaag
     7201 gttattgtag tgtattttat aaagttgatg tacatcgact tattttttaa aacttatatc
     7261 aaaaaaatat gagaaaagta tatactatta aaatgttttt tactataatt ataaatagtt
     7321 aaatatttgg gaattaaata aatatatagg aggcaatttt tagagataca tagcaatgta
     7381 aaggtatcaa atagttattt cataatttat tatgatgaca taattttcag tagatagttg
     7441 taaattaaca ctgaaataga tgtttatatt aagaagactt taatatcata attaatcaaa
     7501 atcttgtgtc aaacatttaa acgaatttta agttgataac aattggcaaa acattgttca
     7561 aacatcacaa tgataaagca tattatcagt attgtagtgt gtggaaaatg acagccatct
     7621 aaggagaaaa atgatgaaaa gaatattggt agtattttta atgttagcaa ttatattggc
     7681 aggttgttct aataaaggtg aaaagtatca aaaagatatt gataaagtgt acaaagaaca
     7741 gaatcaaatg aataaaattg cctcgaaagt acaaaacact attaaaacag acattaaaca
     7801 agaagacagt aatacacatg tttataaaga tggtaaagtc attgttattg gtattcaatt
     7861 atataaagat cgtgaaaaaa tgtattattt cgcatatgaa ataaaagatg gtaaggcaga
     7921 gattaataga gaaatagacc caattaagta tatgaaagac cataaagcag attatgaaga
     7981 tgaagggtct cggatccgat atcgcccgac gcgaggctgg atggccttcc ccattatgat
     8041 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     8101 agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc
     8161 gatcattgga ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg
     8221 gttggcatgg attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg
     8281 tgcatggagc cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc
     8341 accactccaa gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac
     8401 ccttggcaga acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg
     8461 ggcagcgttg ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg
     8521 ctaggctggc ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg
     8581 tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt
     8641 ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg
     8701 atctgcatcg caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag
     8761 cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt
     8821 ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc
     8881 atcctctctc gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag
     8941 gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag
     9001 acattaacgc ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt
     9061 gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat
     9121 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     9181 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     9241 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     9301 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     9361 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     9421 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     9481 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     9541 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     9601 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     9661 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     9721 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     9781 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     9841 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     9901 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     9961 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
    10021 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
    10081 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
    10141 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
    10201 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
    10261 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
    10321 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
    10381 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
    10441 gccccagtgc tgcaatgata ccgcgagatc cacgctcacc ggctccagat ttatcagcaa
    10501 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
    10561 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
    10621 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
    10681 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
    10741 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
    10801 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
    10861 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
    10921 gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag
    10981 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
    11041 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
    11101 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
    11161 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
    11221 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
    11281 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
    11341 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa
//
