LOCUS       V004509                 6913 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V004509
VERSION     V004509
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6913)
  AUTHORS   Kalivoda EJ, Horzempa J, Stella NA, Sadaf A, Kowalski RP, Nau GJ,
            Shanks RM.
  TITLE     New vector tools with a hygromycin resistance marker for use with
            opportunistic pathogens
  JOURNAL   Mol. Biotechnol. 48 (1), 7-14 (2011)
   PUBMED   20972648
REFERENCE   2  (bases 1 to 6913)
  AUTHORS   Kalivoda EJ, Horzempa JE, Sadaf AA, Stella NA, Nau GJ, Shanks RM.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI
            1015, 203 Lothrop St, Pittsburgh, PA 15213, USA
REFERENCE   3  (bases 1 to 6913)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6913)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol.
            Biotechnol."; date: "2011"; volume: "48"; issue: "1"; pages: "7-14"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (19-DEC-2009) Ophthalmology, University of Pittsburgh, EEI 1015, 203
            Lothrop St, Pittsburgh, PA 15213, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            complement(399..508)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
     terminator      complement(929..956)
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1048..1134)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     1484..1500
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(1524..2216)
                     /gene="SCEI"
                     /label="Intron-encoded endonuclease I-SceI"
                     /note="Intron-encoded endonuclease I-SceI from
                     Saccharomyces cerevisiae (strain ATCC 204508 / S288c).
                     Accession#: P03882"
     protein_bind    complement(2246..2262)
                     /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)."
     promoter        complement(2270..2300)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2315..2336)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      2523..2745
                     /label="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2793..3740
                     /label="Rep101(Ts)"
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
     CDS             complement(4031..5026)
                     /gene="hyg"
                     /label="Hygromycin-B 7''-O-kinase"
                     /note="Hygromycin-B 7''-O-kinase from Streptomyces
                     hygroscopicus. Accession#: P09979"
     promoter        complement(5157..5261)
                     /label="AmpR promoter"
     CDS             complement(5361..6161)
                     /label="URA3"
                     /note="orotidine-5'-phosphate decarboxylase, required for
                     uracil biosynthesis"
     promoter        complement(6162..6382)
                     /label="URA3 promoter"
     misc_feature    6410..6913
                     /label="CEN/ARS"
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tctaagaaac cattattatc atgacattaa cctataaaaa aggcctctcg agctagagtc
       61 gatcttcgcc agcagggcga ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc
      121 ggtgagccag agtttcagca ggccgcccag gcggcccagg tcgccattga tgcgggccag
      181 ctcgcggacg tgctcatagt ccacgacgcc cgtgattttg tagccctggc cgacggccag
      241 caggtaggcc gacaggctca tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc
      301 gtctggaagg cagtacacct tgataggtgg gctgcccttc ctggttggct tggtttcatc
      361 agccatccgc ttgccctcat ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg
      421 attcccgttg agcaccgcca ggtgcgaata agggacagtg aagaaggaac acccgctcgc
      481 gggtgggcct acttcaccta tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc
      541 tacacgaacc ctttggcaaa atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa
      601 aatcgctata atgaccccga agcagggtta tgcagcggaa aagcgctgct tccctgctgt
      661 tttgtggaat atctaccgac tggaaacagg caaatgcagg aaattactga actgagggga
      721 caggcgagag acgatgccaa agagctacac cgacgagctg gccgagtggg ttgaatcccg
      781 cgcggccaag aagcgccggc gtgatgaggc tgcggttgcg ttcctggcgg tgagggcgga
      841 tgtcgatatg cgtaaggaga aaataccgca tcaggcgcat atttgaatgt atttagaaaa
      901 ataaacaaaa agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
      961 aatttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     1021 cgcaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     1081 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     1141 ggcagttccc tactctcgca tggggagacc ccacactacc atcggcgcta cggcgtttca
     1201 cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaattct
     1261 gttttatcag accgcttctg cgttctgatt taatctgtat caggctgaaa atcttctctc
     1321 atccgccaaa acagccaagc tcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     1381 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     1441 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     1501 gccaagcttg catgccctta ttatttcagg aaagtttcgg aggagatagt gttcggcagt
     1561 ttgtacatca tctgcgggat caggtacggt ttgatcaggt tgtagaagat caggtaagac
     1621 atagaatcga tgtagatgat cggtttgttt ttgttgattt ttacgtaaca gttcagttgg
     1681 aatttgttac gcagaccctt aaccaggtat tctacttctt cgaaagtgaa agactgggtg
     1741 ttcagtacga tcgatttgtt ggtagagttt ttgttgtaat cccatttacc accatcatcc
     1801 atgaaccagt atgccagaga catcggggtc aggtagtttt caaccaggtt gttcgggatg
     1861 gtttttttgt tgttaacgat gaacaggtta gccagtttgt tgaaagcttg gtgtttgaaa
     1921 gtctgggcgc cccaggtgat taccaggtta cccaggtggt taacacgttc ttttttgtgc
     1981 ggcggggaca gtacccactg atcgtacagc agacatacgt ggtccatgta tgctttgttt
     2041 ttccactcga actgcataca gtaggtttta ccttcatcac gagaacggat gtaagcatca
     2101 cccaggatca gaccgatacc tgcttcgaac tgttcgatgt tcagttcgat cagctgggat
     2161 ttgtattctt tcagcagttt agagttcgga cccaggttca ttacctggtt tttttgatgc
     2221 atatgcgttc tcctcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2281 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2341 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgca
     2401 gcgatttgcc cgagcttgcg agggtgctac ttaagccttt agggttttaa ggtctgtttt
     2461 gtagaggagc aaacagcgtt tgcgacatcc ttttgtaata ctgcggaact gactaaagta
     2521 gtgagttata cacagggctg ggatctattc tttttatctt tttttattct ttctttattc
     2581 tataaattat aaccacttga atataaacaa aaaaaacaca caaaggtcta gcggaattta
     2641 cagagggtct agcagaattt acaagttttc cagcaaaggt ctagcagaat ttacagatac
     2701 ccacaactca aaggaaaagg actagtaatt atcattgact agcccatctc aattggtata
     2761 gtgattaaaa tcacctagac caattgagat gtatgtctga attagttgtt ttcaaagcaa
     2821 atgaactagc gactagtcgc tatgacttaa cggagcatga aaccaagcta attttatgct
     2881 gtgtggcact actcaacccc acgattgaaa accctacaag gaaagaacgg acggtatcgt
     2941 tcacttataa ccaatacgtt cagatgatga acatcagtag ggaaaatgct tatggtgtat
     3001 tagctaaagc aaccagagag ctgatgacga gaactgtgga aatcaggaat cctttggtta
     3061 aaggctttga gattttccag tggacaaact atgccaagtt ctcaagcgaa aaattagaat
     3121 tagtttttag tgaagagata ttgccttatc ttttccagtt aaaaaaattc ataaaatata
     3181 atctggaaca tgttaagtct tttgaaaaca aatactctat gaggatttat gagtggttat
     3241 taaaagaact aacacaaaag aaaactcaca aggcaaatat agagattagc cttgatgaat
     3301 ttaagttcat gttaatgctt gaaaataact accatgagtt taaaaggctt aaccaatggg
     3361 ttttgaaacc aataagtaaa gatttaaaca cttacagcaa tatgaaattg gtggttgata
     3421 agcgaggccg cccgactgat acgttgattt tccaagttga actagataga caaatggatc
     3481 tcgtaaccga acttgagaac aaccagataa aaatgaatgg tgacaaaata ccaacaacca
     3541 ttacatcaga ttcctaccta cataacggac taagaaaaac actacacgat gctttaactg
     3601 caaaaattca gctcaccagt tttgaggcaa aatttttgag tgacatgcaa agtaagtatg
     3661 atctcaatgg ttcgttctca tggctcacgc aaaaacaacg aaccacacta gagaacatac
     3721 tggctaaata cggaaggatc tgaggttctt atggctcttg tatctatcag tgaagcatca
     3781 agactaacaa acaaaagtag aacaactgtt caccgttaca tatcaaaggg aaaactgtcc
     3841 atatgcacag atgaaaacgg tgtaaaaaag atagatacat cagagctttt acgagttttt
     3901 ggtgcaatca atctaaagta tatatgagta aacttggtct gacaatcgat gcgaattggc
     3961 cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa
     4021 cgaattgtca ggcgccgggg gcggtgtccg gcggccccca gaggaactgc gccagttcct
     4081 ccggatcggt gaagccggag agatccagcg gggtctcctc gaacacctcg aagtcgtgca
     4141 ggaaggtgaa ggcgagcagt tcgcgggcga agtcctcggt ccgcttccac tgcgccccgt
     4201 cgagcagcgc ggccaggatc tcgcggtcgc cccggaaggc gttgagatgc agttgcacca
     4261 ggctgtagcg ggagtctccc gcatagacgt cggtgaagtc gacgatcccg gtgacctcgg
     4321 tcgcggccag gtccacgaag atgttggtcc cgtgcaggtc gccgtggacg aaccggggtt
     4381 cgcggccggc cagcagcgtg tccacgtccg gcagccagtc ctccaggcgg tccagcagcc
     4441 ggggcgagag gtagccccac ccgcggtggt cctcgacggt cgccgcgcgg cgttcccgca
     4501 gcagttccgg gaagacctcg gaatgggggg tgagcacggt gttcccggtc agcggcaccc
     4561 tgtgcagccg gccgagcacc cggccgagtt cgcgggccag ggcgagcagc gcgttccggt
     4621 cggtcgtgcc gtccatcgcg gaccgccagg tggtgccggt catccggctc atcaccaggt
     4681 agggccacgg ccaggctccg gtgccgggcc gcagctcgcc gcggccgagg aggcggggca
     4741 ccggcaccgg ggcgtccgcc aggaccgcgt acgcctccga ctccgacgcg aggctctccg
     4801 gaccgcacca gtgctcgccg aacagcttga tcaccgggtc gggctcgccg accagtacgg
     4861 ggttggtgct ctcgccgggc acccgcagca ccggcggcac cggcagcccg agctcctcca
     4921 gggctcggcg ggccagcggc tcccagaatt cctggtcgtt ccgcaggctc gcgtaggaat
     4981 catccgaatc aatacggtcg agaagtaaca gggattcttg tgtcatatga atcttactcc
     5041 tttgttaaat tatttttgtt taagcatttt gaatttgtat gcatcttaga acaatctcaa
     5101 agttccaagc tcttgcaagt ctagatattg agtcaaaaaa aaaattttca actcatactc
     5161 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5221 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5281 ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat acatgcattt
     5341 acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc ttcccagcct
     5401 gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa tagtcctctt
     5461 ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct atactgttga
     5521 cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa ccaatcgtaa
     5581 ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa atctttgtcg
     5641 ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga gcccttgcat
     5701 gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc tgccgcctgc
     5761 ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat gtctgcccat
     5821 tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc aatgtcagca
     5881 aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt tagcggctta
     5941 actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag taaacaaatt
     6001 ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac atccaatgaa
     6061 gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc aacaggacta
     6121 ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct tcgtttcctg
     6181 caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt ttcttcaaca
     6241 ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc ttccttctgt
     6301 tcggagatta ccgaatcaaa aaaatttcaa agaaaccgaa atcaaaaaaa agaataaaaa
     6361 aaaaatgatg aattgaattg aaaagctagc ttatcgatgg gtccttttca tcacgtgcta
     6421 taaaaataat tataatttaa attttttaat ataaatatat aaattaaaaa tagaaagtaa
     6481 aaaaagaaat taaagaaaaa atagtttttg ttttccgaag atgtaaaaga ctctaggggg
     6541 atcgccaaca aatactacct tttatcttgc tcttcctgct ctcaggtatt aatgccgaat
     6601 tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt acattttact
     6661 tatcgttaat cgaatgtata tctatttaat ctgcttttct tgtctaataa atatatatgt
     6721 aaagtacgct ttttgttgaa attttttaaa cctttgttta tttttttttc ttcattccgt
     6781 aactcttcta ccttctttat ttactttcta aaatccaaat acaaaacata aaaataaata
     6841 aacacagagt aaattcccaa attattccat cattaaaaga tacgaggcgc gtgtaagtta
     6901 caggcaagcg atc
//
