LOCUS       62056_17685        7489 bp DNA     circular SYN 10-FEB-2021
DEFINITION  Cloning vector pMYC21, complete sequence.
ACCESSION   MT572318
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7489)
  AUTHORS   Gross S, Schnell B, Haack P, Auerbach D, Mueller R.
  TITLE     In vivo and in vitro reconstitution of unique key steps in 
            cystobactamid antibiotic biosynthesis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7489)
  AUTHORS   Gross S, Schnell B, Haack P, Auerbach D, Mueller R.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JUN-2020) Microbial Natural Products, Helmholtz 
            Institute for Pharmaceutical Research Saarland, Helmholtz Center for
            Infection Research, Saarland University Campus E8.1, Saarbruecken, 
            Saarland 66123, Germany
REFERENCE   3  (bases 1 to 7489)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (04-JUN-2020) Microbial Natural Products, Helmholtz Institute for 
            Pharmaceutical Research Saarland, Helmholtz Center for Infection 
            Research, Saarland University Campus E8.1, Saarbruecken, Saarland 
            66123, Germany"
FEATURES             Location/Qualifiers
     source          1..7489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        28..248
                     /label=URA3 promoter
     CDS             249..1049
                     /label=URA3
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     rep_origin      1166..1711
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     promoter        2079..2181
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2182..2838
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     repeat_region   3052..3080
                     /label=from Himar1
                     /note="from Himar1"
     oriT            3230..3339
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             3372..3740
                     /label=traJ
                     /note="oriT-recognizing protein"
     misc_feature    3812..4315
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
     misc_feature    4593..6421
                     /label=mx9 int building block
                     /note="mx9 int building block"
     gene            4770..6416
                     /gene="int"
                     /label=int
     CDS             4770..6416
                     /codon_start=1
                     /transl_table=11
                     /gene="int"
                     /product="Int"
                     /label=int
                     /note="site specific recombinase"
                     /protein_id="QRG42414.1"
                     /translation="MALRGASDATTNPSRLVQSVAAGPRATPWGVSASWYLLGRTATGE
                     YIVSSDAAKKGHPMATAAERLPTSPIDVNALALEVARLVALQQQSATPPSSGRTFGAVA
                     DDWLITEAKRLVCPDNERRHLRHMEALWGMTDVELTPRVVKAHLAGLLKPEGPLSAATV
                     NKVRSTGKRIIKAAQINGEWGPVNPFGVLDREKEAKAERLTLTAAECRAVLPHFRADRR
                     REFLFQVFLGPRPGEEKALLKEDVDVEARTVIFRRSNGRDTTKTGRERRVPVPDELWPV
                     LLDAMQASPSDLVFPNAKGERQRADTKMTRVLRTALSAAGVVVGWDYICRTQGCGYRDV
                     QSGGARQERRCPACDKRMWASGRPKPAVWYGLRHTAATLHRKAGCDPLVIKLVLGHAAV
                     DTTDDVYTHLDEDYCRAELNKLSLKAPPPPPTHQGGSDGGPDSGRNTYGEGGTMHGLGD
                     LQHHRARAWEARALPTELPPRNLAGGIPAPLLSVKDVAASLSVSTAKVYQLLAAGVLPT
                     VWVGQSRRVKREDLDAYIARATATGGKRGGK"
     misc_feature    5560..5591
                     /gene="int"
                     /label=attP
                     /note="attP"
     regulatory      6473..6546
                     /label=kanR promoter
                     /note="kanR promoter"
                     /regulatory_class="promoter"
     CDS             6547..7359
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      7440..7489
                     /label=tD2 terminator
                     /note="tD2 terminator"
                     /regulatory_class="terminator"
ORIGIN
        1 gttaactaat gcgctagccc acagcttttc aattcaattc atcatttttt ttttattctt
       61 ttttttgatt tcggtttctt tgaaattttt ttgattcggt aatctccgaa cagaaggaag
      121 aacgaaggaa ggagcacaga cttagattgg tatatatacg catatctagt gttgaagaaa
      181 catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa
      241 gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg
      301 ctgccaagct atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg
      361 ttcgtaccac caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac
      421 taaaaacaca tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa
      481 aggcattatc cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg
      541 gtaatacagt caaattgcag tactctgcgg gtgtctacag aatagcagaa tgggcagaca
      601 ttacgaatgc acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag
      661 aagaagtaac aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct
      721 ccctatctac tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt
      781 ttgttatcgg ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt
      841 tgattatgac acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata
      901 gaaccgtgga tgatgttgtc tctacaggat ctgacattat tattgttgga agaggactat
      961 ttgcaaaggg aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag
     1021 catatttgag aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgtc
     1081 tactaaactc acaaattaga gcttcaattt aattatatca gttattaccc gtgcacgtcc
     1141 agctcgagtt aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct
     1201 ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact
     1261 ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt
     1321 agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct
     1381 gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag
     1441 gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc
     1501 tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac
     1561 accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc
     1621 ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg
     1681 atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc
     1741 ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc
     1801 gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc
     1861 ctgtatcaca tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca
     1921 cttcactgac accctcatca gtgccaacat agtaagccag catacactcc gctaccgctg
     1981 atacctgtga cggaagatca cttcgcagaa taaataaatc ctggtgtccc tgttgatacc
     2041 gggaagccct gggccaactt ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc
     2101 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
     2161 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
     2221 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
     2281 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
     2341 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
     2401 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
     2461 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
     2521 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
     2581 ttccctaaag ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc
     2641 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
     2701 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
     2761 gccgtctgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
     2821 gagtggcagg gcggggcgta atttttttaa ggcagttatt ggtgccctta aacgcctggt
     2881 gctacgcctg aataagtgat aataagcgga tgaatggcag aaattcgaaa gcaaattcga
     2941 cccggtcgtc ggttcagggc agggtcgtta aatagccgct tatgtctatt gctggtttac
     3001 cggtttattg actaccggaa gcagtgtgac cgtgtgcttc ttagatgtgt cagaccgggg
     3061 acttatcagc caacctgtta gcagaacttt agcgcttttc cgctgcataa ccctgcttcg
     3121 gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat acaggatttt
     3181 gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg ggcaggatag
     3241 gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat tcgcacctgg
     3301 cggtgctcaa cgggaatcct gctctgcgag gctggccggc taccgccggg gtaacagatg
     3361 agggcaagcg gatggctgat gaaaccaagc caaccaggaa gggcagccca cctatcaagg
     3421 tgtactgcct tccagacgaa cgaagagcga ttgaggaaaa ggcggcggcg gccggcatga
     3481 gcctgtcggc ctacctgctg gccgtcggcc agggctacaa aatcacgggc gtcgtggact
     3541 atgagcacgt ccgcgagctg gcccgcatca atggcgacct gggccgcctg ggcggcctgc
     3601 tgaaactctg gctcaccgac gacccgcgca cggcgcggtt cggtgatgcc acgatcctcg
     3661 ccctgctggc gaagatcgaa gagaagcagg acgagcttgg caaggtcatg atgggcgtgg
     3721 tccgcccgag ggcagagcca tgactttttt agccgctaaa acggccgggg ggtgcgcgtg
     3781 agcggccgcg taacgtacgt aggtcctttt catcacgtgc tataaaaata attataattt
     3841 aaatttttta atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa
     3901 aaatagtttt tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac
     3961 cttttatctt gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg
     4021 tagaagacca cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta
     4081 tatctattta atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg
     4141 aaatttttta aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt
     4201 atttactttc taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc
     4261 aaattattcc atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc
     4321 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4381 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     4441 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     4501 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4561 gtggaccata gtttaaacgc attattaatt aagcggcaat cccttatgtc gcaatttcgt
     4621 ggctcactcc aagagtgaaa acccgtcact cgcgtgtatc ggatttatcg cgcatgtgtt
     4681 gtttttcatg cgggatgcga gcgttgcgtc aagtaaaatc gcatccgata tgttgttttc
     4741 agggattcgt tcggttaccg tactgattag tggcgctcag gggtgcgtcg gacgccacta
     4801 ccaacccctc tcgacttgtg cagtccgtcg ccgccggccc gcgtgcgact ccgtggggtg
     4861 tcagtgcgtc gtggtacctg ctagggcgta cagcaacggg ggagtacatc gtgagtagcg
     4921 acgcggcgaa gaagggccat ccaatggcaa ctgcggcgga gcggttgccg acgtcaccaa
     4981 tcgacgtcaa cgctctggcg ctggaggtgg cccggcttgt ggccctccag cagcaaagtg
     5041 cgacgccgcc atcgtccggc cgcactttcg gcgcggtggc ggatgactgg ctcatcactg
     5101 aggccaagcg cctcgtgtgc cccgacaatg agcgccgcca tcttcgccac atggaggcgc
     5161 tctggggcat gacggatgtg gagctgacgc cgcgcgtcgt gaaggcgcac ctggcgggac
     5221 ttctcaagcc agaggggccg ctgagcgcag ccaccgtcaa taaggtgcgc tctaccggca
     5281 agcgcatcat caaggcggcg caaatcaacg gcgagtgggg cccggtgaat cctttcggcg
     5341 tgctcgaccg cgaaaaagag gcgaaggccg agcgcctcac gctgacggca gcggagtgcc
     5401 gggcggtgct cccgcacttc cgcgcggacc ggcgccgcga gtttctcttc caggtctttc
     5461 tggggccacg ccccggcgaa gagaaggcgc tcctcaagga agatgtggac gtcgaggcgc
     5521 gcaccgtcat tttccggcgc agcaatggac gagacacgac aaagacggga cgcgagcgtc
     5581 gcgtgccggt gccggatgag ttgtggcccg tgctcctcga tgcgatgcag gccagtccgt
     5641 ctgacctcgt tttcccgaac gcgaagggtg agaggcagcg cgcagacacg aagatgacgc
     5701 gcgtgctgcg cactgcgcta tccgcggctg gtgtcgtggt gggctgggat tacatctgcc
     5761 gcacgcaggg ctgcggctac cgagatgtgc agtctggtgg cgcgcgccag gagcgtcggt
     5821 gccccgcctg cgacaagcgc atgtgggcca gtggtcgccc caaacccgcc gtctggtacg
     5881 ggctccgtca caccgcggcg acactgcaca ggaaggcggg ctgcgacccg ctcgtcatca
     5941 agctcgtgct ggggcatgcg gctgtcgaca ccacggacga cgtgtacacg cacctcgacg
     6001 aggactactg ccgcgccgaa cttaacaagt tgtcgctgaa ggccccgccg ccaccaccta
     6061 ctcaccaggg aggaagtgac ggcggccctg actcaggacg caacacctac ggtgaaggag
     6121 gcaccatgca cggattggga gatttgcagc atcaccgggc gagagcttgg gaagctcgtg
     6181 ctctaccaac tgagctacca ccgcggaact tggccggggg gataccggcg ccgctgctga
     6241 gcgtcaagga cgttgcggct tcactctcag tgagcacggc gaaggtgtac cagctcctcg
     6301 ccgccggtgt cctgcctacc gtgtgggtgg gccagtcgcg ccgcgtcaag cgtgaggacc
     6361 tggacgccta catcgcccgc gcgacggcca ccggcgggaa gcggggtggc aaatgagctg
     6421 agtatacgtc gtaactagtg ccacgttgtg tctcaaaatc tctgatgtta cattgcacaa
     6481 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     6541 ggtgttatga gccatattca acgggaaacg tcttgctcaa ggccgcgatt aaattccaac
     6601 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     6661 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     6721 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     6781 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     6841 actgcgatcc cagggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     6901 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     6961 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     7021 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     7081 tggaaagaaa tgcataagct tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     7141 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     7201 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     7261 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     7321 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     7381 ttgtaacact ggcagagcat tacgctgact tgacgggacc ccgggaccgg tcgccggcgc
     7441 ctcgctagaa gcccagcaat ccctccgggg gttgctgggc tttcttttt
//
