LOCUS       40924_4184        5786 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pAJ63a, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5786)
  AUTHORS   Peters B, Junker A, Brauer K, Muhlthaler B, Kostner D, Mientus M, 
            Liebl W, Ehrenreich A.
  TITLE     Deletion of pyruvate decarboxylase by a new method for efficient 
            markerless gene deletions in Gluconobacter oxydans
  JOURNAL   Appl. Microbiol. Biotechnol. 97 (6), 2521-2530 (2013)
  PUBMED    22940799
REFERENCE   2  (bases 1 to 5786)
  AUTHORS   Peters B, Junker A, Brauer K, Kostner D, Mientus M, Liebl W, 
            Ehrenreich A.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2012) Department of Microbiology, Technische 
            Universitaet Muenchen, Emil-Ramann-Str. 4, Freising, Bayern 85354, 
            Germany
REFERENCE   3  (bases 1 to 5786)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5786)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Microbiol. Biotechnol."; date: "2013"; volume: "97"; issue: "6"; 
            pages: "2521-2530"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-MAY-2012) Department of Microbiology, Technische Universitaet 
            Muenchen, Emil-Ramann-Str. 4, Freising, Bayern 85354, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             401..1192
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1888..2526
                     /codon_start=1
                     /gene="upp"
                     /product="UPRTase"
                     /EC_number="2.4.2.9"
                     /label=upp
                     /note="uracil phosphoribosyltransferase; derived from 
                     Gluconobacter oxydans 621H"
                     /protein_id="AFP65781.1"
                     /translation="MSQALPPIVLDHPLVRHKLTRLRDHNTSTAGFRRLTRELSLLLAY
                     EATRNLSLAPRHIEAPSGPMEGEELDGKKLCFVSILRAGNGLLDGMLDLVPSARVGHIG
                     LRRDHETLEVSEYYFNMPSDVPGRTCIVLDPMLATGHSAAAALTRVKAAGATNPIFACL
                     LAVPEGIAHMAELHPDVQIVTCSIDSHLDEHGYIVPGLGDAGDRLFGTR"
     gene            1888..2526
                     /gene="upp"
                     /label=upp
     oriT            complement(3529..3638)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        complement(3873..3963)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     promoter        complement(4157..4247)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      4607..5195
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5483..5504
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5519..5549
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5557..5573
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5581..5597
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    5606..5662
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5666..5682)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ccttcccaac agttgcgcag cctgaatggc gaatggcgat aagctagctt cacgctgccg
       61 caagcactca gggcgcaagg gctgctaaag gaagcggaac acgtagaaag ccagtccgca
      121 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
      181 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
      241 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
      301 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
      361 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
      421 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
      481 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
      541 ttttgtcaag accgacctgt ccggtgccct gaatgaactc caagacgagg cagcgcggct
      601 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
      661 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
      721 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
      781 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
      841 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
      901 agccgaactg ttcgccaggc tcaaggcgcg gatgcccgac ggcgaggatc tcgtcgtgac
      961 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1021 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1081 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1141 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1201 actctggggt tcggatccgg gcttatcgac tgcacggtgc accaatgctt ctggcgtcag
     1261 gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa atcactgcat aattcgtgtc
     1321 gctcaaggcg cactcccgtt ctggataatg ttttttgcgc cgacatcata acggttctgg
     1381 caaatattca tgacagctta agatgcgtga caaaatacgg ccgcgctttc atacatcgtc
     1441 agaagacgcc tttcagattt tccggagaac cagcatgacc cagagttgcg gcccgaacgg
     1501 gcagaccggt ggcggttgcg gtgttggcct gacaccggaa cagcggcgta tccttcacga
     1561 acacggaacg gaacgacccg gctccagtcc cctcaatgat gagaagcgcg cggggattta
     1621 tgcctgtgcg gcctgcgggc gcccgctttt tgcttcggaa acgaaatatg atagcggcag
     1681 tggctggccc tcgttcttcc aggctctgcc ggatggcgtc gaaaccacgc aggacagccg
     1741 ctatggcatg gtccgcaccg aagtccactg cgcgaactgc aagggccatc tggggcacgt
     1801 ctttcccgat ggtccgccgc ctacgggact tcgttactgc atgaacggtc tggcgctcga
     1861 tttccgcccg gccgaaggag aaaaagcatg agtcaggcgc tgccgcctat cgtcctcgac
     1921 cacccgctgg tgcgccacaa gctgacccgt ctgcgtgacc ataatacgtc cacagccggt
     1981 ttccgccgcc tgacgcgcga actgagcctg ctgctggcct acgaagccac gcgcaatctg
     2041 tcgctggcgc cgcggcatat cgaggcgccg agcggcccga tggaaggcga ggaactggac
     2101 ggcaagaagc tgtgtttcgt ttccatcctg cgcgccggta atggtctgct ggatggcatg
     2161 ctggatctgg tgccgtccgc gcgtgtcggc catatcggtc tgcgccgcga tcacgagacg
     2221 ctggaagtca gcgaatacta tttcaacatg cccagcgatg ttccgggacg gacctgcatc
     2281 gtgctcgatc cgatgctggc aacagggcac tccgccgctg ctgcgctgac gcgcgtgaag
     2341 gctgccggtg cgacgaaccc gatcttcgcc tgcctgctgg cagttcccga gggcattgcc
     2401 cacatggcgg aactgcatcc ggatgtgcag atcgtgacct gttcgatcga cagtcatctg
     2461 gacgagcacg gctatatcgt tccgggcctc ggcgatgcgg gtgaccgtct gttcggtacg
     2521 cgctgaggct ttggcacacg ctcttgtgtg ctgaaacttc cgcggttttg tggcgggtat
     2581 ggaggctcat ccagaaggga gcgttccaat atttcgcgcc actggcggaa gcaacgcgta
     2641 aactcgaccc gacgcgtccg atcacctgcg tcaatgtaat gttctgcgac gctcacaccg
     2701 ataccatcag cgatctcttt gatgtgctgt gcctgaaccg ttattacgga tggtatgtcc
     2761 aaagcggcga tttggaaacg gcagagaagg tactggaaaa agaacttctg gcctggcagg
     2821 agaaactgca tcagccgatt atcatcaccg aatacggcgt ggatacgtta gccgggctgc
     2881 actcaatgta caccgacatg tggagtgaag agtatcagtg tgcatggctg gatatgtatc
     2941 accgcgtctt tgatcgcgtc agcgccgtcg tcggtgaaca ggtatggaat ttcgccgatt
     3001 ttgcgacctc gcaaggcata ttgcgcgttg gcggtaacaa gaaagggatc ttcactcgcg
     3061 accgcaaacc gaagtcggcg gcttttctgc tgcaaaaacg ctggactggc atgaacttcg
     3121 gtgaaaaacc gcagcaggga ggcaaacaat gaatcaacaa ctctcctggc gcaccatcgt
     3181 cggctacagc ctcgggaatt gctaccgcga aatgaccgac caagcgacgc ccaacctgcc
     3241 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3301 ccgggacgcc ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc
     3361 cgacggccag caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg
     3421 ctcttcgttc gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct
     3481 tggtttcatc agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc
     3541 gcagagcagg attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac
     3601 acccgctcgc gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc
     3661 aaggaaagtc tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat
     3721 ataccgaaaa aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct
     3781 tccctgctgt tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga
     3841 actgagggga caggcgagag acgatgccaa agagctcctg aaaatctcga taactcaaaa
     3901 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     3961 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     4021 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg
     4081 tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt
     4141 ggcttctgtt tctatcagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg
     4201 atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc
     4261 gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg
     4321 cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca
     4381 acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc
     4441 agggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaaaa
     4501 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4561 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4621 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4681 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4741 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4801 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4861 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4921 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4981 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5041 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5101 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5161 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5221 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5281 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5341 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5401 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5461 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     5521 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5581 caggaaacag ctatgacatg attacgaatt cgagctcggt acccggggat cctctagagt
     5641 cgacctgcag gcatgcaagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     5701 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     5761 atagcgaaga ggcccgcacc gatcgc
//
