LOCUS       62056_17680        7890 bp DNA     circular SYN 10-FEB-2021
DEFINITION  Cloning vector pMYC20, complete sequence.
ACCESSION   MT572317
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7890)
  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  JOURNAL   Unpublished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 7890)
  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 7890)
  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            "Unpublished"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..7890
                     /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..6423
                     /label=mx8 int building block
                     /note="mx8 int building block"
     gene            4699..6423
                     /gene="int"
                     /label=int
     CDS             4699..6423
                     /codon_start=1
                     /transl_table=11
                     /gene="int"
                     /product="integrase"
                     /label=int
                     /protein_id="QRG42409.1"
                     /translation="MVASRSRSWRRFSRRRGRRLRPKRRPTRRQRPSWRGRQGGAVGNV
                     YRKKATRYLWMWWFDAAGARQYASTGEEDEVKARAVLKATEKRVAAEKDQGLGASGPVT
                     VEAYAAKWMEGRVTLRSVMDDRSRMQHHVLPSLGDKELGVLTRRHVLALVREVRAKGLA
                     PRTVRHVYALVRTMLSDAVVDGLLERSPCDLRERRGEIPAIRDADPRWRDSAVYSRAEV
                     VALITDERIPLERRVLYAVAFVGAVRIGEASALQVRDWDGTVNPLGKLWVTKTFNHKVG
                     EVYETTKTDTPRQMPVHPWLSTMLASWLAYGWARMYARTPRQDDLILPSPTGMHQRPQG
                     VLRRLHADLDLLGFRRRRTHDARRTFISLARADGADKYLLQWVTHGRRSDMVDAYTTPP
                     WDALCREVAKLKLELPKQEEETMACRVPSCDSGVTADAGKEKPQQVRELAGAGVARSTG
                     LEPVLLGSFYANPRQLPTKSEGLDAESAKLARTGAVDWHKPVTAPHPLAVVRQQMGVLK
                     SGAGRERKVLASQLVDASLMALERLAAALHRTHAADDRWSDCPACRAVADGESPSAT"
     misc_feature    6055..6083
                     /gene="int"
                     /label=attP site
                     /note="attP site"
     promoter        6456..6484
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             6532..7719
                     /label=TcR
                     /note="tetracycline efflux protein"
     regulatory      7841..7890
                     /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 aaaagacgga cgaggtgctc actcagactg
     4621 aggcggcagc cctcgcaagt gtcagcacat ccaccattcg caactggcag gcgcagggac
     4681 tcctgacggc tgggcatcgt ggtcgcgtca agaagtcgga gttggaggcg tttctcgcgt
     4741 cgccgcgggc gaaggctacg cccgaagagg aggccgacgc gaaggcagcg gccatcctgg
     4801 cgcggaaggc agggcggcgc cgtgggtaac gtctatcgca agaaggccac tcgctacttg
     4861 tggatgtggt ggttcgacgc tgcgggcgcg cggcagtacg cgagcacggg cgaagaagac
     4921 gaagtgaagg cgcgcgccgt cctgaaggcg accgagaagc gcgtggccgc ggagaaggac
     4981 cagggactcg gcgcgtccgg tcccgtcacc gtcgaggcct acgccgccaa gtggatggag
     5041 gggcgcgtca cgctgcgctc ggtcatggat gaccgctccc ggatgcagca ccacgtgctg
     5101 ccctcgttgg gcgacaagga actcggtgtg ctgacgcgtc ggcacgtgct cgcactggtg
     5161 cgcgaagtgc gcgcaaaggg cctggcgccg cgaacggtgc gccacgtcta cgccctggtg
     5221 cggacgatgc tgagtgacgc ggtggtggac gggctgctgg agcgctcgcc ctgtgacttg
     5281 cgcgagcgcc gcggagagat tccggccatt cgggatgctg accctcgatg gcgggactcg
     5341 gccgtctact cgcgggccga ggtcgtcgca ctgattacgg atgagcgcat cccccttgag
     5401 cggcgcgtgc tctacgccgt ggccttcgtc ggagcggtgc gcatcgggga ggcgtcagcc
     5461 ctgcaggtcc gggactggga cggcaccgtc aaccccttgg ggaagctctg ggtgacgaag
     5521 accttcaacc acaaggtagg agaggtctac gagacgacga agacggacac gccgcggcag
     5581 atgccagtcc acccctggtt gagcacgatg cttgccagtt ggctcgccta cggctgggcg
     5641 cggatgtacg cccggacgcc gcgccaggat gacctgattc tcccgagccc caccgggatg
     5701 caccagcggc cccagggagt cctccgtcgc ctgcatgccg acctggacct cctggggttc
     5761 aggcggagac ggactcacga tgctcgccgg acgttcatca gcctggctcg tgccgacggt
     5821 gccgacaaat acctactcca gtgggtcacc cacggcaggc gctccgacat ggtcgacgcc
     5881 tacacgacgc cgccgtggga tgcgctctgt agggaagtgg cgaagttgaa gctcgaactg
     5941 cccaagcagg aggaggaaac catggcctgc cgagtgccaa gctgtgacag cggtgtgaca
     6001 gcggacgctg gaaaagaaaa accccagcaa gtccgagaac ttgctggggc aggggtggcg
     6061 aggagtacgg gacttgaacc cgtactcctg ggctccttct acgccaatcc gcgccagttg
     6121 ccaacaaaat ctgagggctt agacgcggag tccgccaaac tggcacggac tggcgcagtt
     6181 gactggcaca agcccgtgac agcgccccat ccgcttgccg tggtgcgtca gcagatgggg
     6241 gtcctcaagt ccggcgccgg acgtgagcgc aaggtgctgg cgtcccaact cgtggacgcc
     6301 tccctgatgg ccctggagcg gcttgccgcg gccctgcacc gcacccatgc cgccgatgac
     6361 cgatggtccg actgcccggc ctgccgcgcc gtggcggacg gcgagagccc ttccgctacc
     6421 tgagtatacg tcgtaactag tattaattct catgtttgac agcttatcat cgattagctt
     6481 taatgcggta gtttatcaca gttaaattgc taacgcagtc aggcaccgtg tatgaaatct
     6541 aacaatgcgc tcatcgtcat cctcggcacc gtcaccctgg atgctgtagg cataggcttg
     6601 gttatgccgg tactgccggg cctcttgcgg gacattgtcc attccgacag catcgccagt
     6661 cactatggcg tgctgcttgc gctatatgcg ttgatgcaat ttctatgcgc acccgttctc
     6721 ggagcactgt ccgaccgctt tggccgccgc ccagtcctgc tcgcttcgct acttggagcc
     6781 actatcgact acgcgatcat ggcgaccaca cccgtcctgt ggattctcta cgccggacgc
     6841 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
     6901 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
     6961 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttaca tgcaccattc
     7021 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
     7081 tcgcataagg gagagcgccg acccatgccc ttgagagcct tcaacccagt cagctccttc
     7141 cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa
     7201 ctcgtaggac aggtgccggc agcgctctgg gtcattttcg gcgaggaccg ctttcgctgg
     7261 agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa tcttgcacgc cctcgctcaa
     7321 gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga agcaggccat tatcgccggc
     7381 atggcggccg acgcgctggg ctacgtcttg ctggcgttcg cgacgcgagg ctggatggcc
     7441 ttccccatta tgattcttct cgcttccggc ggcatcggga tgcccgcgtt gcaggccatg
     7501 ctgtccaggc aggtagatga cgaccatcag ggacagcttc aaggatcgct cgcggctctt
     7561 accagcctaa cttcgatcat tggaccgctg atcgtcacgg cgatttatgc cgcctcggcg
     7621 agcacatgga acgggttggc atggattgta ggcgccgccc tataccttgt ctgcctcccc
     7681 gcgttgcgtc gcggtgcatg gagccgggcc acctcgacct gaatggaagc cggcggcacc
     7741 tcgctaacgg attcaccact ccaagaattg gagccaatca attcttgcgg agaactgtga
     7801 atgcgcaaac caacccttgg cccgggaccg gtcgccggcg cctcgctaga agcccagcaa
     7861 tccctccggg ggttgctggg ctttcttttt
//
