LOCUS       sequence-271251-        4922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Low Phosphate inducible SBS (bonryl-diphosphate) synthase with 
            C-terminal mCherry).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4922)
  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 4922)
  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-yibDp-SBS-mCherry
FEATURES             Location/Qualifiers
     source          1..4922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..104
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      120..147
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      2897..2943
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3026..3042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3026..3042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3039..3061)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(3222..3767)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3302..3321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3875..3969)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3990..4649)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4564..4583
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4650..4752)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     4839..4861
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4853..4870
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4854..4870
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cagtccagtt acgctggagt ctgcccaggc atcaaataaa acgaaaggct cagtcgaaag
       61 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
      121 gctcaccttc gggtgggcct ttctgcgttt atacacagct aacaccacgt cgtccctatc
      181 tgctgcccta ggtctatgag tggttgctgg ataacgtgcg taattgtgct gatctcttat
      241 atagctgctc tcattatctc tctaccctga agtgactctc tcacctgtaa aaataatatc
      301 tcacaggctt aatagtttct taatacaaag cctgtaaaac gtcaggataa cttctatatt
      361 cagggagacc acaacggttt ccctctacaa ataattttgt ttaactttcg tgtgtaggag
      421 gataatctat gatggaggct caccagattc gccgttcagg caattaccaa ccggcattgt
      481 gggactccaa ttatattcaa tcccttaata caccgtatac ggaagaacgc catcttgacc
      541 gcaaagccga acttatcgtt caagttcgta tcctgctgaa agaaaagatg gagccagtac
      601 aacaattgga gttaattcat gatcttaagt acttaggcct gagtgatttt ttccaagacg
      661 aaatcaagga aatccttggg gtgatctata acgaacataa atgttttcat aataatgagg
      721 tggaaaaaat ggatttatac tttactgcgt tgggctttcg ccttcttcgc cagcacggat
      781 ttaacatttc acaggatgtg tttaactgtt tcaaaaatga aaaggggatc gattttaagg
      841 cttccttagc tcaagatacc aaaggtatgt tgcaattata tgaagcaagc ttcctgttgc
      901 gcaaggggga agatacctta gagttagctc gtgaatttgc tactaaatgc ttacagaaaa
      961 aattggacga agggggcaac gagatcgacg agaacctttt gttatggatt cgccactctc
     1021 ttgatcttcc tctgcattgg cgcatccaat cagtcgaagc acgctggttt atcgatgcgt
     1081 acgcacgccg cccggacatg aatccattga ttttcgagtt ggcaaagttg aactttaaca
     1141 ttatccaagc tactcatcag caggaattga aagatctttc tcgttggtgg agtcgcttat
     1201 gcttcccaga aaagttaccc ttcgtgcgcg accgtttggt cgagtccttc ttttgggcgg
     1261 tcgggatgtt cgagccgcac cagcacggat accagcgcaa gatggcagcc acaattattg
     1321 ttttggcgac agtcattgac gatatctatg atgtatacgg gacactggat gagttggaat
     1381 tgttcacaga cactttcaaa cgctgggata ctgagagcat cacacgcctt ccctattata
     1441 tgcaactgtg ttattgggga gttcacaact atatctcgga cgccgcctac gacattctta
     1501 aagaacacgg gtttttttgt ttgcagtact tacgcaaatc agtagtagac ctggttgaag
     1561 cctactttca tgaggctaag tggtatcatt cgggttacac gccgagcctt gatgaatact
     1621 taaatattgc taagatcagc gtagcttctc cagcaattat cagtcctaca tatttcacgt
     1681 tcgccaacgc aagtcacgac accgccgtaa tcgactcctt gtatcagtac cacgacattc
     1741 tttgtttggc agggatcatt ttgcgtcttc cggacgattt ggggacaagt tactttgagc
     1801 ttgcgcgcgg cgacgtgcca aagactattc aatgctacat gaaggaaaca aatgcttccg
     1861 aggaagaagc agtcgaacat gtgaagtttt tgattcgcga ggcgtggaaa gacatgaata
     1921 cagcaattgc agcgggatac ccttttcccg atggcatggt cgcgggagct gcaaacatcg
     1981 ggcgtgtagc ccagttcatt tacttgcacg gagatggatt tggggtgcag cacagtaaga
     2041 cctacgaaca cattgcaggt ttgttgttcg agccatatgc ggggggttca ggcgggggct
     2101 cgggtggcat ggcaattatc aaggaattta tgcgtttcaa agttcacatg gaggggtctg
     2161 ttaacggaca cgaattcgaa attgagggag agggggaggg tcgtccctac gaagggacgc
     2221 aaaccgccaa acttaaggtc acgaaaggtg ggccgttacc tttcgcctgg gacattctgt
     2281 ccccgcaatt tatgtacggc tcaaaagcat atgtgaagca tcccgccgat attcccgatt
     2341 accttaaatt atccttccct gaaggattca aatgggagcg tgtcatgaac ttcgaagatg
     2401 ggggtgttgt cacggtaacc caggattcat cgttacaaga tggtgagttc atttacaaag
     2461 ttaagttgcg cggaacaaac tttccctcgg acggccctgt aatgcagaag aagacaatgg
     2521 gatgggaggc gtcaagtgaa cgtatgtacc ccgaagatgg agcacttaag ggggaaatca
     2581 agcagcgctt aaagctgaag gacggagggc attatgatgc cgaggtgaaa actacatata
     2641 aagcaaagaa acctgtacaa cttccgggag cttacaatgt taacatcaag ttagacatta
     2701 cttcacacaa cgaagactac acaatcgtcg agcagtacga acgcgcggaa ggccgtcaca
     2761 gcaccggggg catggatgag ctgtataaat aatgataaac tagttgactc gagtctggta
     2821 aaactagcat tcgacctagc ataaccccgc ggggcctctt cgggggtctc gcggggtttt
     2881 ttgctgaaag aagcttcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2941 tatactgacc atttaaatca tacctgacca ggctaggtgg aggctcagtg atgataagtc
     3001 tgcgatggtg gatgcatgtg tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3061 tcagagggca caatcctatt ccgcgctatc cgacaatctc caagacatta ggtggagttc
     3121 agttcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
     3181 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
     3241 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
     3301 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
     3361 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
     3421 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
     3481 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
     3541 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
     3601 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     3661 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     3721 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     3781 ttaagtctga cgctctattc aacaaagccg ccgtccatgg gtagggggct tcaaatcgtc
     3841 cccccatacg atataagttg ttactagtgc ttggattctc accaataaaa aacgcccggc
     3901 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
     3961 acaggagtcc aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac
     4021 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc
     4081 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
     4141 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     4201 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     4261 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     4321 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     4381 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga
     4441 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     4501 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     4561 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     4621 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat
     4681 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     4741 acgtgccgat caacgtctca ttgatacctg aaacaaaacc catcgtacgg ccaaggaagt
     4801 ctccaataac tgtgatccac cacaagcgcc agggttttcc cagtcacgac gttgtaaaac
     4861 gacggccagt catgcataat ccgcacgcat ctggaataag gaagtgccat tccgcctgac
     4921 ct
//
