LOCUS       40924_47318        4419 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pXS99A, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4419)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Tools for genetic engineering in the amino acid-producing bacterium 
            Corynebacterium glutamicum
  JOURNAL   J. Biotechnol. 104 (1-3), 287-299 (2003)
  PUBMED    12948646
REFERENCE   2  (bases 1 to 4419)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JAN-2003) Department of Genetics, University of 
            Bielefeld, Universitaetsstrasse 25, Bielefeld D-33615, Germany
REFERENCE   3  (bases 1 to 4419)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4419)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. 
            Biotechnol. 104 (1-3), 287-299 (2003)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JAN-2003) Department of Genetics, University of Bielefeld, 
            Universitaetsstrasse 25, Bielefeld D-33615, Germany"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4419
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      260..346
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      438..465
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     CDS             886..1653
                     /codon_start=1
                     /gene="aad9"
                     /product="Aad9"
                     /label=aad9
                     /note="spectinomycin adenylyltransferase"
                     /protein_id="AAO65210.1"
                     /translation="MRRIYLNTYEQINKVKKILRKHLKNNLIGTYMFGSGVESGLKPNS
                     DLDFLVVVSEPLTDQSKEILIQKIRPISKKIGDKSNLRYIELTIIIQQEMVPWNHPPKQ
                     EFIYGEWLQELYEQGYIPQKELNSDLTIMLYQAKRKNKRIYGNYDLEELLPDIPFSDVR
                     RAIMDSSEELIDNYQDDETNSILTLCRMILTMDTGKIIPKDIAGNAVAESSPLEHRERI
                     LLAVRSYLGENIEWTNENVNLTINYLNNRLKKL"
     gene            886..1653
                     /gene="aad9"
                     /label=aad9
     rep_origin      1851..2439
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2625..2765)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        2951..3028
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             3029..4108
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        4343..4372
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    4380..4396
                     /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)."
ORIGIN
        1 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
       61 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
      121 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
      181 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga
      241 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      301 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
      361 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
      421 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
      481 tctttttgtt tatttttcta aatacattca aatatgtatc cgctcatgcc ctctggtaag
      541 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
      601 aggggatcaa gatctatcga ttttcgttcg tgaatacatg ttataataac tataactaat
      661 aacgtaacgt gactggcaag agatattttt aaaacaatga ataggtttac acttacttta
      721 gttttatgga aatgaaagat catatcatat ataatctaga ataaaattaa ctaaaataat
      781 tattatctag ataaaaaatt tagaagccaa tgaaatctat aaataaacta aattaagttt
      841 atttaattaa caactatgga tataaaatag gtactaatca aaatagtgag gaggatatat
      901 ttgaatacat acgaacaaat taataaagtg aaaaaaatac ttcggaaaca tttaaaaaat
      961 aaccttattg gtacttacat gtttggatca ggagttgaga gtggactaaa accaaatagt
     1021 gatcttgact ttttagtcgt cgtatctgaa ccattgacag atcaaagtaa agaaatactt
     1081 atacaaaaaa ttagacctat ttcaaaaaaa ataggagata aaagcaactt acgatatatt
     1141 gaattaacaa ttattattca gcaagaaatg gtaccgtgga atcatcctcc caaacaagaa
     1201 tttatttatg gagaatggtt acaagagctt tatgaacaag gatacattcc tcagaaggaa
     1261 ttaaattcag atttaaccat aatgctttac caagcaaaac gaaaaaataa aagaatatac
     1321 ggaaattatg acttagagga attactacct gatattccat tttctgatgt gagaagagcc
     1381 attatggatt cgtcagagga attaatagat aattatcagg atgatgaaac caactctata
     1441 ttaactttat gccgtatgat tttaactatg gacacgggta aaatcatacc aaaagatatt
     1501 gcgggaaatg cagtggctga atcttctcca ttagaacata gggagagaat tttgttagca
     1561 gttcgtagtt atcttggaga gaatattgaa tggactaatg aaaatgtaaa tttaactata
     1621 aactatttaa ataacagatt aaaaaaatta taaaaaaatt gaaaaaatgg tggaaacact
     1681 tttttcaatt tttttgtttt attatttaat atttgggaaa tattcattct aattggtaat
     1741 cagattttag aaaacaataa acccttgcat atgatcatga ccaaaatccc ttaacgtgag
     1801 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1861 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1921 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1981 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2041 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2101 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2161 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2221 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2281 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2341 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2401 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2461 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2521 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2581 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     2641 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     2701 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     2761 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2821 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2881 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg
     2941 catttacgtt gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
     3001 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     3061 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     3121 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     3181 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     3241 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     3301 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     3361 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     3421 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     3481 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     3541 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     3601 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     3661 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     3721 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     3781 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     3841 gggatacgac gataccgaag acagctcatg ttatatcccg ccgtcaacca ccatcaaaca
     3901 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     3961 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     4021 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     4081 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagcgcg
     4141 aattgatctg gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca
     4201 ggcagccatc ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt
     4261 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
     4321 gcaaatattc tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat
     4381 tgtgagcgga taacaatttc acacaggaaa cagaccatg
//
