LOCUS       Exported                2783 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Low phosphate inducible yibD gene promoter expressing GFPuv 
            reporter.
ACCESSION   .
VERSION     .
KEYWORDS    pHCKan-yibDp-GFPuv
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2783)
  AUTHORS   Menacho-Melgar R, Ye Z, Moreb EA, Yang T, Efromson JP, Decker JS, 
            Wang R, Lynch MD
  TITLE     Scalable, two-stage, autoinduction of recombinant protein expression
            in E. coli utilizing phosphate depletion.
  JOURNAL   Biotechnol Bioeng. 2020 May 22. doi: 10.1002/bit.27440.
  PUBMED    32441815
REFERENCE   2  (bases 1 to 2783)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2783
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             216..935
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(249..277)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     872..892
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      1122..1153
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1154..1256
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1257..2072
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1330..1349)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2093..2120
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2132..2719)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2212..2231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      2741..2770
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cagtccagtt acgctggagt ctgaggctcg tcctgaatga tatcaagctt gaattcgttg
       61 tgcgtaattg tgctgatctc ttatatagct gctctcatta tctctctacc ctgaagtgac
      121 tctctcacct gtaaaaataa tatctcacag gcttaatagt ttcttaatac aaagcctgta
      181 aaacgtcagg ataacttctt gtaggaggat aatctatggc tagcaaagga gaagaacttt
      241 tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt
      301 ctgtcagtgg agagggtgaa ggtgatgcta catacggaaa gcttaccctt aaatttattt
      361 gcactactgg aaaactacct gttccatggc caacacttgt cactactttc tcttatggtg
      421 ttcaatgctt ttcccgttat ccggatcata tgaaacggca tgactttttc aagagtgcca
      481 tgcccgaagg ttatgtacag gaacgcacta tatctttcaa agatgacggg aactacaaga
      541 cgcgtgctga agtcaagttt gaaggtgata cccttgttaa tcgtatcgag ttaaaaggta
      601 ttgattttaa agaagatgga aacattctcg gacacaaact cgagtacaac tataactcac
      661 acaatgtata catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaattc
      721 gccacaacat tgaagatgga tccgttcaac tagcagacca ttatcaacaa aatactccaa
      781 ttggcgatgg ccctgtcctt ttaccagaca accattacct gtcgacacaa tctgcccttt
      841 cgaaagatcc caacgaaaag cgtgaccaca tggtccttct tgagtttgta actgctgctg
      901 ggattacaca tggcatggat gagctctaca aataatgagg atccccggct tatcggtcag
      961 tttcacctga tttacgtaaa aacccgcttc ggcgggtttt tgcttttgga ggggcagaaa
     1021 gatgaatgac tgtccacgac gctataccca aaagaaagac gaattctcta gatatcgctc
     1081 aatactgacc atttaaatca tacctgacct ccatagcaga aagtcaaaag cctccgaccg
     1141 gaggcttttg acttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
     1201 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga
     1261 gccatattca acgggaaacg tcttgctcga ggccgcgatt aaattccaac atggatgctg
     1321 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
     1381 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
     1441 ccaatgatgt tacagatgag atggtcaggc taaactggct gacggaattt atgcctcttc
     1501 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
     1561 cagggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1621 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1681 acggcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1741 gtgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     1801 tgcataagct tttgccattc tcaccggatt cagtcgtcac ccatggtgat ttctcacttg
     1861 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     1921 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     1981 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     2041 agtttcactt gatgctcgat gagtttttct gagggcccaa atgtaatcac ctggctcacc
     2101 ttcgggtggg cctttctgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2161 atcacaaaaa tcgatgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2221 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2281 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2341 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2401 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2461 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2521 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2581 ttggtatctg cgctctgctg aagccagtta cctcggaaaa agagttggta gctcttgatc
     2641 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2701 cagaaaaaaa ggatctcaag aagatccttt gattttctac cgaagaaagg cccacccgtg
     2761 aaggtgagcc agtgagttga ttg
//
