LOCUS       40924_25536        6248 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pING1001, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6248)
  AUTHORS   Ghosh IN, Landick R.
  TITLE     OptSSeq: High-throughput sequencing readout of growth enrichment 
            defines optimal gene expression elements for homoethanologenesis
  JOURNAL   ACS Synth Biol (2016) In press
  PUBMED    27404024
REFERENCE   2  (bases 1 to 6248)
  AUTHORS   Ghosh IN, Landick RC.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUL-2016) DOE Great Lakes Bioenergy Research Center, 
            University of Wisconsin-Madison, 1552 University Avenue, Madison, WI
            53726, United States
REFERENCE   3  (bases 1 to 6248)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6248)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JUL-2016) DOE Great Lakes Bioenergy Research Center, University 
            of Wisconsin-Madison, 1552 University Avenue, Madison, WI 53726, 
            United States"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    144..160
                     /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)."
     protein_bind    177..193
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             240..953
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     regulatory      1013..1064
                     /label=Phage P22 terminator
                     /note="Phage P22 terminator"
                     /regulatory_class="terminator"
     regulatory      1201..1248
                     /label=B. subtilis glnA terminator
                     /note="B. subtilis glnA terminator"
                     /regulatory_class="terminator"
     protein_bind    complement(1260..1279)
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     regulatory      1307..1335
                     /label=Promoter#1
                     /note="Promoter#1"
                     /regulatory_class="promoter"
     CDS             1391..2065
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     terminator      2113..2184
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2200..2227
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2365..3024)
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(3025..3794)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     promoter        5172..5263
                     /label=AmpR promoter
     CDS             5264..6052
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
ORIGIN
        1 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
       61 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc acctcgagaa
      121 aatttatcaa aaagagtgtt gacttgtgag cggataacaa tgatacttag attcaattgt
      181 gagcggataa caatttcaca catctagaat tagtttaact ttaagaagga gatatacata
      241 tgagcaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      301 atgttaatgg gcacaaattt tctgtccgtg gagagggtga aggtgatgct acaaacggaa
      361 aactcaccct taaatttatt tgcactactg gaaaactacc tgttccgtgg ccaacacttg
      421 tcactactct gacctatggt gttcaatgct tttcccgtta tccggatcac atgaaacggc
      481 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaacgcact atatctttca
      541 aagatgacgg gacctacaag acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta
      601 atcgtatcga gttaaagggt attgatttta aagaagatgg aaacattctt ggacacaaac
      661 tcgagtacaa ctttaactca cacaatgtat acatcacggc agacaaacaa aagaatggaa
      721 tcaaagctaa cttcaaaatt cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc
      781 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
      841 tgtcgacaca atctgtcctt tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc
      901 ttgagtttgt aactgctgct gggattacac atggcatgga tgagctctac aaactcgagt
      961 aagcttgata tcgaattcct gcagcccggg ggatccctag ttctagaaat aagttaaata
     1021 aagccctgag tttaaccgct cggggctttt tgcgttttaa gcacactaga gcggccatgc
     1081 atcccagagc ctgataaaaa cggtgaatcc gttagcgagg tgccgccggc ttcgcgcgtt
     1141 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtaact
     1201 actcaatccc ttggcactaa aagtgtcagg ggatttttta tgttaataca ccatcacaga
     1261 attgtgagcg ctcacaatct aggtctatga gtggttgctg gacctcytka yaattaatca
     1321 tccggctcgb ataatgbgtg gaattggtag agtaggcacc catcaccacc acactagcag
     1381 gaggtaaaac atggcttcct ccgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg
     1441 tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc
     1501 gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc
     1561 ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc
     1621 tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat
     1681 gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga
     1741 gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc cggttatgca
     1801 gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct
     1861 gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt
     1921 taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat
     1981 caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc
     2041 tgaaggtcgt cactccaccg gtgcttaata acgctgatag tgctagtgta gatcgctact
     2101 agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2161 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     2221 ttctgcgcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     2281 cagacaaggt atagggcggc gaggcggcta cagccgatag tctggaacag cgcacttacg
     2341 ggttgctgcg caacccaagt gctaccggcg cggcagcgtg acccgtgtcg gcggctccaa
     2401 cggctcgcca tcgtccagaa aacacggctc atcgggcatc ggcaggcgct gctgcccgcg
     2461 ccgttcccat tcctccgttt cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc
     2521 gggctggctg ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc ccggatacag
     2581 ggtcgggatg cggcgcaggt cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc
     2641 aaccaccacg gcggcactga acaccgacag gcgcaactgg tcgcggggct ggccccacgc
     2701 cacgcggtca ttgaccacgt aggccgacac ggtgccgggg ccgttgagct tcacgacgga
     2761 gatccagcgc tcggccacca agtccttgac tgcgtattgg accgtccgca aagaacgtcc
     2821 gatgagcttg gaaagtgtct tctggctgac caccacggcg ttctggtggc ccatctgcgc
     2881 cacgaggtga tgcagcagca ttgccgccgt gggtttcctc gcaataagcc cggcccacgc
     2941 ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc
     3001 gatttctctg gactgcgtgg ccatgcttat ctccatgcgg tagggtgccg cacggttgcg
     3061 gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg cgttgcgtaa
     3121 aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc ggggggtgcc
     3181 gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc tttcgcattt
     3241 cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc
     3301 cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc
     3361 gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca ctcgccgccg
     3421 tgaggctcgg ggggcaggcg ggcgggcttc gccttcgact gcccccactc gcataggctt
     3481 gggtcgttcc aggcgcgtca aggccaagcc gctgcgcggt cgctgcgcga gccttgaccc
     3541 gccttccact tggtgtccaa ccggcaagcg aagcgcgcag gccgcaggcc ggaggctttt
     3601 ccccagagaa aattaaaaaa attgatgggg caaggccgca ggccgcgcag ttggagccgg
     3661 tgggtatgtg gtcgaaggct gggtagccgg tgggcaatcc ctgtggtcaa gctcgtgggc
     3721 aggcgcagcc tgtccatcag cttgtccagc agggttgtcc acgggccgag cgaagcgagc
     3781 cagccggtgg ccgctcgcgg ccatcgtcca catatccacg ggctggcaag ggagcgcagc
     3841 gaccgcgcag ggcgaagccc ggagagcaag cccgtagggc gccgcagccg ccgtaggcgg
     3901 tcacgacttt gcgaagcaaa gtctagtgag tatactcaag cattgagtgg cccgccggag
     3961 gcaccgcctt gcgctgcccc cgtcgagccg gttggacacc aaaagggagg ggcaggcatg
     4021 gcggcatacg cgatcatgcg atgcaagaag ctggcgaaaa tgggcaacgt ggcggccagt
     4081 ctcaagcacg cctaccgcga gcgcgagacg cccaacgctg acgccagcag gacgccagag
     4141 aacgagcact gggcggccag cagcaccgat gaagcgatgg gccgactgcg cgagttgctg
     4201 ccagagaagc ggcgcaagga cgctgtgttg gcggtcgagt acgtcatgac ggccagcccg
     4261 gaatggtgga agtcggccag ccaagaacag caggcggcgt tcttcgagaa ggcgcacaag
     4321 tggctggcgg acaagtacgg ggcggatcgc atcgtgacgg ccagcatcca ccgtgacgaa
     4381 accagcccgc acatgaccgc gttcgtggtg ccgctgacgc aggacggcag gctgtcggcc
     4441 aaggagttca tcggcaacaa agcgcagatg acccgcgacc agaccacgtt tgcggccgct
     4501 gtggccgatc tagggctgca acggggcatc gagggcagca aggcacgtca cacgcgcatt
     4561 caggcgttct acgaggccct ggagcggcca ccagtgggcc acgtcaccat cagcccgcaa
     4621 gcggtcgagc cacgcgccta tgcaccgcag ggattggccg aaaagctggg aatctcaaag
     4681 cgcgttgaga cgccggaagc cgtggccgac cggctgacaa aagcggttcg gcaggggtat
     4741 gagcctgccc tacaggccgc cgcaggagcg cgtgagatgc gcaagaaggc cgatcaagcc
     4801 caagagacgg cccgagacct tcgggagcgc ctgaagcccg ttctggacgc cctggggccg
     4861 ttgaatcggg atatgcaggc caaggccgcc gcgatcatca aggccgtggg cgaaaagctg
     4921 ctgacggaac agcgggaagt ccagcgccag aaacaggccc agcgccagca ggaacgcggg
     4981 cgcgcacatt tccccgaaaa gtgccacctg gcggcgttgt gacaatttac cgaacaactc
     5041 cgcggccggg aagccgatct cggcttgaac gaattcctgc ttatccacgg aatctgtggg
     5101 taactttgta tgtgtccgca gcgcccgccg cagtctcacg cccggagcgt agcgaccgag
     5161 tgagctagct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     5221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgaggg aagcggtgat
     5281 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     5341 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     5401 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     5461 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     5521 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     5581 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     5641 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     5701 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     5761 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     5821 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     5881 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     5941 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     6001 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataatgtct
     6061 aacaattcgt tcaagccgag gggccgcaag atccggccac gatgacccgg tcgtcggttc
     6121 agggcagggt cgttaaatag ccgcttatgt ctattgctgg tttaccggtt tattgactac
     6181 cggaagcagt gtgaccgtgt gcttcttact acgtcagtgt cgtgccatag atctcataag
     6241 ggagagcg
//
