LOCUS       sequence-489556-        8191 bp    DNA     circular SYN 16-OCT-2025
DEFINITION  pCS27 containing aroL, ppsA, tktA, aroG(fbr) from E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8191)
  AUTHORS   Shen X, Wang J, Wang J, Chen Z, Yuan Q, Yan Y
  TITLE     High-level De novo biosynthesis of arbutin in engineered Escherichia
            coli.
  JOURNAL   Metab Eng. 2017 Jul;42:52-58. doi: 10.1016/j.ymben.2017.06.001. Epub
            2017 Jun 2.
  PUBMED    28583673
REFERENCE   2  (bases 1 to 8191)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 16, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Metab 
            Eng."; date: "2017-07"; volume: "42:52-58. doi"; pages: " 
            10.1016/j.ymben.2017.06.001. Epub 2017 Jun 2"
COMMENT     Sequence Label: pCS-APTA
FEATURES             Location/Qualifiers
     source          1..8191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5936..6022
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(6057..6074)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6191..6736)
                     /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(6271..6290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6850..6944)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6975..7769)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7086..7105)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7696..7715
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7948..7966
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    7989..8005
                     /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)."
     RBS             8047..8058
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 tgacggtcgc ggagatcgtc gaaagggaag agtgggcggg atttcgcgcc agagaaacgg
       61 cggcgctgga agcggtaact gcgccatcca ccgttatcgc tacaggcggc ggcattattc
      121 tgacggaatt taatcgtcac ttcatgcaaa ataacgggat cgtggtttat ttgtgtgcgc
      181 cagtatcagt cctggttaac cgactgcaag ctgcaccgga agaagattta cggccaacct
      241 taacgggaaa accgctgagc gaagaagttc aggaagtgct ggaagaacgc gatgcgctat
      301 atcgcgaagt tgcgcatatt atcatcgacg caacaaacga acccagccag gtgatttctg
      361 aaattcgcag cgccctggca cagacgatca attgttgaca tatgaggaga tataccatgt
      421 ccaacaatgg ctcgtcaccg ctggtgcttt ggtataacca actcggcatg aatgatgtag
      481 acagggttgg gggcaaaaat gcctccctgg gtgaaatgat tactaatctt tccggaatgg
      541 gtgtttccgt tccgaatggt ttcgccacaa ccgccgacgc gtttaaccag tttctggacc
      601 aaagcggcgt aaaccagcgc atttatgaac tgctggataa aacggatatt gacgatgtta
      661 ctcagcttgc gaaagcgggc gcgcaaatcc gccagtggat tatcgacact cccttccagc
      721 ctgagctgga aaacgccatc cgcgaagcct atgcacagct ttccgccgat gacgaaaacg
      781 cctcttttgc ggtgcgctcc tccgccaccg cagaagatat gccggacgct tcttttgccg
      841 gtcagcagga aaccttcctc aacgttcagg gttttgacgc cgttctcgtg gcagtgaaac
      901 atgtatttgc ttctctgttt aacgatcgcg ccatctctta tcgtgtgcac cagggttacg
      961 atcaccgtgg tgtggcgctc tccgccggtg ttcaacggat ggtgcgctct gacctcgcat
     1021 catctggcgt gatgttctcc attgataccg aatccggctt tgaccaggtg gtgtttatca
     1081 cttccgcatg gggccttggt gagatggtcg tgcagggtgc ggttaacccg gatgagtttt
     1141 acgtgcataa accgacactg gcggcgaatc gcccggctat cgtgcgccgc accatggggt
     1201 cgaaaaaaat ccgcatggtt tacgcgccga cccaggagca cggcaagcag gttaaaatcg
     1261 aagacgtacc gcaggaacag cgtgacatct tctcgctgac caacgaagaa gtgcaggaac
     1321 tggcaaaaca ggccgtacaa attgagaaac actacggtcg cccgatggat attgagtggg
     1381 cgaaagatgg ccacaccggt aaactgttca ttgtgcaggc gcgtccggaa accgtgcgct
     1441 cacgcggtca ggtcatggag cgttatacgc tgcattcaca gggtaagatt atcgccgaag
     1501 gccgtgctat cggtcatcgc atcggtgcgg gtccggtgaa agtcatccat gacatcagcg
     1561 aaatgaaccg catcgaacct ggcgacgtgc tggttactga catgaccgac ccggactggg
     1621 aaccgatcat gaagaaagca tctgccatcg tcaccaaccg tggcggtcgt acctgtcacg
     1681 cggcgatcat cgctcgtgaa ctgggcattc cggcggtagt gggctgtgga gatgcaacag
     1741 aacggatgaa agacggtgag aacgtcactg tttcttgtgc cgaaggtgat accggttacg
     1801 tctatgcgga gttgctggaa tttagcgtga aaagctccag cgtagaaacg atgccggatc
     1861 tgccgttgaa agtgatgatg aacgtcggta acccggaccg tgctttcgac ttcgcctgcc
     1921 taccgaacga aggcgtgggc cttgcgcgtc tggaatttat catcaaccgt atgattggcg
     1981 tccacccacg cgcactgctt gagtttgacg atcaggaacc gcagttgcaa aacgaaatcc
     2041 gcgagatgat gaaaggtttt gattctccgc gtgaatttta cgttggtcgt ctgactgaag
     2101 ggatcgcgac gctgggtgcc gcgttttatc cgaagcgcgt cattgtccgt ctctctgatt
     2161 ttaaatcgaa cgaatatgcc aacctggtcg gtggtgagcg ttacgagcca gatgaagaga
     2221 acccgatgct cggcttccgt ggcgcgggcc gctatgtttc cgacagcttc cgcgactgtt
     2281 tcgcgctgga gtgtgaagca gtgaaacgtg tgcgcaacga catgggactg accaacgttg
     2341 agatcatgat cccgttcgtg cgtaccgtag atcaggcgaa agcggtggtt gaagaactgg
     2401 cgcgtcaggg gctgaaacgt ggcgagaacg ggctgaaaat catcatgatg tgtgaaatcc
     2461 cgtccaacgc cttgctggcc gagcagttcc tcgaatattt cgacggcttc tcaattggct
     2521 caaacgatat gacgcagctg gcgctcggtc tggaccgtga ctccggcgtg gtgtctgaat
     2581 tgttcgatga gcgcaacgat gcggtgaaag cactgctgtc gatggctatc cgtgccgcga
     2641 agaaacaggg caaatatgtc gggatttgcg gtcagggtcc gtccgaccac gaagactttg
     2701 ccgcatggtt gatggaagag gggatcgata gcctgtctct gaacccggac accgtggtgc
     2761 aaacctggtt aagcctggct gaactgaaga aataagtcga gaggagatat accatgtcct
     2821 cacgtaaaga gcttgccaat gctattcgtg cgctgagcat ggacgcagta cagaaagcca
     2881 aatccggtca cccgggtgcc cctatgggta tggctgacat tgccgaagtc ctgtggcgtg
     2941 atttcctgaa acacaacccg cagaatccgt cctgggctga ccgtgaccgc ttcgtgctgt
     3001 ccaacggcca cggctccatg ctgatctaca gcctgctgca cctcaccggt tacgatctgc
     3061 cgatggaaga actgaaaaac ttccgtcagc tgcactctaa aactccgggt cacccggaag
     3121 tgggttacac cgctggtgtg gaaaccacca ccggtccgct gggtcagggt attgccaacg
     3181 cagtcggtat ggcgattgca gaaaaaacgc tggcggcgca gtttaaccgt ccgggccacg
     3241 acattgtcga ccactacacc tacgccttca tgggcgacgg ctgcatgatg gaaggcatct
     3301 cccacgaagt ttgctctctg gcgggtacgc tgaagctggg taaactgatt gcattctacg
     3361 atgacaacgg tatttctatc gatggtcacg ttgaaggctg gttcaccgac gacaccgcaa
     3421 tgcgtttcga agcttacggc tggcacgtta ttcgcgacat cgacggtcat gacgcggcat
     3481 ctatcaaacg cgcagtagaa gaagcgcgcg cagtgactga caaaccttcc ctgctgatgt
     3541 gcaaaaccat catcggtttc ggttccccga acaaagccgg tacccacgac tcccacggtg
     3601 cgccgctggg cgacgctgaa attgccctga cccgcgaaca actgggctgg aaatatgcgc
     3661 cgttcgaaat cccgtctgaa atctatgctc agtgggatgc gaaagaagca ggccaggcga
     3721 aagaatccgc atggaacgag aaattcgctg cttacgcgaa agcttatccg caggaagccg
     3781 ctgaatttac ccgccgtatg aaaggcgaaa tgccgtctga cttcgacgct aaagcgaaag
     3841 agttcatcgc taaactgcag gctaatccgg cgaaaatcgc cagccgtaaa gcgtctcaga
     3901 atgctatcga agcgttcggt ccgctgttgc cggaattcct cggcggttct gctgacctgg
     3961 cgccgtctaa cctgaccctg tggtctggtt ctaaagcaat caacgaagat gctgcgggta
     4021 actacatcca ctacggtgtt cgcgagttcg gtatgaccgc gattgctaac ggtatctccc
     4081 tgcacggtgg cttcctgccg tacacctcca ccttcctgat gttcgtggaa tacgcacgta
     4141 acgccgtacg tatggctgcg ctgatgaaac agcgtcaggt gatggtttac acccacgact
     4201 ccatcggtct gggcgaagac ggcccgactc accagccggt tgagcaggtc gcttctctgc
     4261 gcgtaacccc gaacatgtct acatggcgtc cgtgtgacca ggttgaatcc gcggtcgcgt
     4321 ggaaatacgg tgttgagcgt caggacggcc cgaccgcact gatcctctcc cgtcagaacc
     4381 tggcgcagca ggaacgaact gaagagcaac tggcaaacat cgcgcgcggt ggttatgtgc
     4441 tgaaagactg cgccggtcag ccggaactga ttttcatcgc taccggttca gaagttgaac
     4501 tggctgttgc tgcctacgaa aaactgactg ccgaaggcgt gaaagcgcgc gtggtgtcca
     4561 tgccgtctac cgacgcattt gacaagcagg atgctgctta ccgtgaatcc gtactgccga
     4621 aagcggttac tgcacgcgtt gctgtagaag cgggtattgc tgactactgg tacaagtatg
     4681 ttggcctgaa cggtgctatc gtcggtatga ccaccttcgg tgaatctgct ccggcagagc
     4741 tgctgtttga agagttcggc ttcactgttg ataacgttgt tgcgaaagca aaagaactgc
     4801 tgtaagcatg caggagatat accatgaatt atcagaacga cgatttacgc atcaaagaaa
     4861 tcaaagagtt acttcctcct gtcgcattgc tggaaaaatt ccccgctact gaaaatgccg
     4921 cgaatacggt tgcccatgcc cgaaaagcga tccataagat cctgaaaggt aatgatgatc
     4981 gcctgttggt tgtgattggc ccatgctcaa ttcatgatcc tgtcgcggca aaagagtatg
     5041 ccactcgctt gctggcgctg cgtgaagagc tgaaagatga gctggaaatc gtaatgcgcg
     5101 tctattttga aaagccgcgt accacggtgg gctggaaagg gctgattaac gatccgcata
     5161 tggataatag cttccagatc aacgacggtc tgcgtatagc ccgtaaattg ctgcttgata
     5221 ttaacgacag cggtctgcca gcggcaggtg agtttctcaa catgatcacc ccacaatatc
     5281 tcgctgacct gatgagctgg ggcgcaattg gcgcacgtac caccgaatcg caggtgcacc
     5341 gcgaactggc atcagggctt tcttgtccgg tcggcttcaa aaatggcacc gacggtacga
     5401 ttaaagtggc tatcgatgcc attaatgccg ccggtgcgcc gcactgcttc ctgtccgtaa
     5461 cgaaatgggg gcattcggcg attgtgaata ccagcggtaa cggcgattgc catatcattc
     5521 tgcgcggcgg taaagagcct aactacagcg cgaagcacgt tgctgaagtg aaagaagggc
     5581 tgaacaaagc aggcctgcca gcacaggtga tgatcgattt cagccatgct aactcgtcca
     5641 aacaattcaa aaagcagatg gatgtttgtg ctgacgtttg ccagcagatt gccggtggcg
     5701 aaaaggccat tattggcgtg atggtggaaa gccatctggt ggaaggcaat cagagcctcg
     5761 agagcgggga gccgctggcc tacggtaaga gcatcaccga tgcctgcatc ggctgggaag
     5821 ataccgatgc tctgttacgt caactggcga atgcagtaaa agcgcgtcgc gggtaaaagc
     5881 ttgatatcga attcctgcag cccgggggat cccatggtac gcgtgctaga ggcatcaaat
     5941 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     6001 cgctctcctg agtaggacaa atccgccgcc ctagacctag ggatatattc cgcttcctcg
     6061 ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg
     6121 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
     6181 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
     6241 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
     6301 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
     6361 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     6421 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     6481 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     6541 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     6601 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     6661 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     6721 accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct agatttcagt
     6781 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgttact
     6841 agtgcttgga ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc
     6901 cagatggagt tctgaggtca ttactggatc tatcaacagg agtccaagcg agctctcgaa
     6961 ccccagagtc ccgctcagaa gaactcgtca agaaggcgat agaaggcgat gcgctgcgaa
     7021 tcgggagcgg cgataccgta aagcacgagg aagcggtcag cccattcgcc gccaagctct
     7081 tcagcaatat cacgggtagc caacgctatg tcctgatagc ggtccgccac acccagccgg
     7141 ccacagtcga tgaatccaga aaagcggcca ttttccacca tgatattcgg caagcaggca
     7201 tcgccatggg tcacgacgag atcctcgccg tcgggcatgc gcgccttgag cctggcgaac
     7261 agttcggctg gcgcgagccc ctgatgctct tcgtccagat catcctgatc gacaagaccg
     7321 gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg cttggtggtc gaatgggcag
     7381 gtagccggat caagcgtatg cagccgccgc attgcatcag ccatgatgga tactttctcg
     7441 gcaggagcaa ggtgagatga caggagatcc tgccccggca cttcgcccaa tagcagccag
     7501 tcccttcccg cttcagtgac aacgtcgagc acagctgcgc aaggaacgcc cgtcgtggcc
     7561 agccacgata gccgcgctgc ctcgtcctgc agttcattca gggcaccgga caggtcggtc
     7621 ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga acacggcggc atcagagcag
     7681 ccgattgtct gttgtgccca gtcatagccg aatagcctct ccacccaagc ggccggagaa
     7741 cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc ctcatcctgt ctcttgatca
     7801 gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg
     7861 cagggcttcc caaccttacc agagggcgcc ccagctggca attccgacgt ctaagaaacc
     7921 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
     7981 ctcgagaatt gtgagcggat aacaagatac tgagcacatc agcaggacgc actgaccgaa
     8041 ttcattaaag aggagaaagg taccatgaca caacctcttt ttctgatcgg gcctcggggc
     8101 tgtggtaaaa caacggtcgg aatggccctt gccgattcgc ttaaccgtcg gtttgtcgat
     8161 accgatcagt ggttgcaatc acagctcaat a
//
