LOCUS       sequence-271204-        2997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 based expression of human glycine acyltransferase with an 
            N-terminal chitin binding domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2997)
  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 2997)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol 
            Bioeng. 2020 May 22. doi: 10.1002/bit.27440."
COMMENT     Sequence Label: pHCKan-T7-CBD-hGLY
FEATURES             Location/Qualifiers
     source          1..2997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      65..96
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        97..199
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             200..1015
                     /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(273..292)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1036..1063
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1075..1662)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1155..1174)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1684..1713)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     1793..1812
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1793..1811
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1843..1865
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     RBS             1873..1884
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1894..2046
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNVP
                     ALWQLQ"
     CDS             2974..2991
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gacgaattct ctagatatcg ctcaatactg accatttaaa tcatacctga cctccatagc
       61 agaaagtcaa aagcctccga ccggaggctt ttgacttgat cggcacgtaa gaggttccaa
      121 ctttcaccat aatgaaataa gatcactacc gggcgtattt tttgagttat cgagattttc
      181 aggagctaag gaagctaaaa tgagccatat tcaacgggaa acgtcttgct cgaggccgcg
      241 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
      301 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
      361 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca ggctaaactg
      421 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
      481 atggttactc accactgcga tcccagggaa aacagcattc caggtattag aagaatatcc
      541 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
      601 tcctgtttgt aattgtcctt ttaacggcga tcgcgtattt cgtctcgctc aggcgcaatc
      661 acgaatgaat aacggtttgg ttggtgcgag tgattttgat gacgagcgta atggctggcc
      721 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
      781 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
      841 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
      901 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
      961 taatcctgat atgaataaat tgcagtttca cttgatgctc gatgagtttt tctgagggcc
     1021 caaatgtaat cacctggctc accttcgggt gggcctttct gcgttgctgg cgtttttcca
     1081 taggctccgc ccccctgacg agcatcacaa aaatcgatgc tcaagtcaga ggtggcgaaa
     1141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1501 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttacctcgga
     1561 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     1621 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgattttc
     1681 taccgaagaa aggcccaccc gtgaaggtga gccagtgagt tgattgcagt ccagttacgc
     1741 tggagtctga ggctcgtcct gaatgatatc aagcttgaat tcgttcattg cataatacga
     1801 ctcactatag ggagaccaca acggtttccc actagaaata attttgttta actttaagaa
     1861 ggagatatac ataaagagga gaaatactag atgacaaatc ctggtgtaag tgcctggcaa
     1921 gttaataccg catataccgc tgggcagtta gtcacttata acggcaagac ctacaagtgc
     1981 ttgcagcctc acacatcctt ggcaggttgg gaaccgtcca atgtacccgc cctttggcaa
     2041 cttcagggct ctgccggtag tgcggcgggt tccggtgaat ttatgctggt tctgcacaac
     2101 agccaaaaac tgcaaattct gtacaaatct ctggaaaaat ctattcctga aagcattaaa
     2161 gtttatggtg cgatcttcaa catcaaggat aaaaaccctt tcaatatgga ggtgctggtc
     2221 gacgcgtggc ctgattacca aatcgtcatc acccgtccgc agaagcaaga aatgaaagat
     2281 gaccaggacc actatactaa cacctaccac atcttcacta aagcgccgga caaactggaa
     2341 gaagttctga gctactccaa tgttatctcc tgggaacaaa cgctgcagat tcaaggttgc
     2401 caggaaggtc tggatgaggc gattcgtaag gtcgctacct ctaagtccgt tcaagtcgat
     2461 tacatgaaaa ccatcctgtt tatcccggag ctgccgaaga aacacaaaac ctcctctaac
     2521 gacaagatgg agctgttcga agtagacgat gacaacaaag agggtaactt ttccaatatg
     2581 ttcctggacg cctcccacgc cggtctggtt aatgagcact gggcgttcgg taaaaacgaa
     2641 cgctccctga agtacattga gcgttgtctg caggattttc tgggttttgg tgtcctgggt
     2701 ccagagggtc aactggtctc ttggatcgtt atggaacagt cttgtgaact gcgtatgggt
     2761 tatactgtgc caaaataccg tcaccaaggt aatatgctgc aaatcggtta tcatctggag
     2821 aagtacctga gccagaagga aatcccgttc tatttccatg ttgccgataa taacgaaaag
     2881 agcctgcaag ccctgaacaa tctgggcttc aagatctgcc cttgcggttg gcaccagtgg
     2941 aagtgcacgc ctaaaaagta ctgtggcggt ggccatcatc accatcacca ttaatga
//
