LOCUS       sequence-358709-        6092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CcCDP in E. coli BL21(DE3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6092)
  AUTHORS   Schwaiger KN, Voit A, Wiltschi B, Nidetzky B
  TITLE     Engineering cascade biocatalysis in whole cells for bottom-up 
            synthesis of cello-oligosaccharides: flux control over three 
            enzymatic steps enables soluble production.
  JOURNAL   Microb Cell Fact. 2022 Apr 9;21(1):61. doi: 
            10.1186/s12934-022-01781-w.
  PUBMED    35397553
REFERENCE   2  (bases 1 to 6092)
  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: "Microb Cell
            Fact."; date: "2022-04-9"; volume: "21(1):61. doi"; pages: " 
            10.1186/s12934-022-01781-w"
COMMENT     Sequence Label: CcCDP_p15A
FEATURES             Location/Qualifiers
     source          1..6092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             38..60
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             80..97
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3065..3112
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3167..3961)
                     /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(3278..3297)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3888..3907
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     RBS             3968..3979
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(3989..5071)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     5033..5052
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5072..5149)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(5286..5303)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5420..5964)
                     /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(5500..5519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6072..6091
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6072..6090
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(6091..6092,1..23)
                     /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)."
ORIGIN
        1 aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga
       61 tatacatatg agaggatcgc atcaccatca ccatcacgga tccattacca aagtgacccc
      121 gcgtagctgc aaactgaccc cgttcgagat tctgaaacaa aaattcggta gcattattgc
      181 gatgagcggc tgcgcggatg cgatctacag cgacagcagc tgcaagaacg tgatcggtat
      241 taccggcctg ccgatcaaag cgccggtgat tcaggttctg ctggagaaca gcaagctggc
      301 ggcgttcctg aacgaatacg tgcaaaacaa aagcgtttgc ctggcgctgg agggtatgcg
      361 ttattgcctg gtgtttgaag acggccagaa ggttaaactg ccgcaagagc tggacgatat
      421 cattcagagc accgtgagcg atgttaaaaa ctgggcgggt accctgaccg agaacggcga
      481 acacgtgatc gacctgaaga aaccgagccc gggtccgcac ttctacgtta acctgctgat
      541 tggtaaccgt ctgggcttta accgtaccct gcaaaccacc ccgaagagcg tggttgatcg
      601 tttcggtcgt ggcagctttc gtagccatgc ggcgacccag gttctggcga cccgttacga
      661 catgcgtcaa gaggaaaacg gcttcccggc gaaccgtcag ttttacctgt atgagaacgg
      721 caagcaaatc ttctacagcg gcagcgtgga cgatgacaac atcctggaag cgaaatgcat
      781 tcacagctgc aaccgtaccg ttattaagta taaaaccgcg tgcaagctgg agatcacccg
      841 taccattttt ctgtgcccgc agaagaaagg tctgccgctg gcgaccgaga tccaaatcat
      901 caacatcaag aacaacagcg aacgtaaacg taacctgagc atcacctaca ccggcatgtt
      961 tggtaccgcg gcgacccacg cgatctttga agatgtgacc tacaccaacg ttattatgca
     1021 gagcgcggcg ctgtataacg acaacggcga gttcatcggc attaccccgg attactatcg
     1081 taagaaatac aacgaagaca cccgtttcgt gaccatgttt gttcgtaagg acggtaaacg
     1141 tatcttcccg aagagcttct gcaccgatta taacgacttt gtgggtaacg gtaccctgga
     1201 gcacccggcg ggtggcgaac acctgaacaa caagctgaac cgtaaaggtc cgggcttctt
     1261 tgcgctgggt accccgttca ccattatgcc gggccaagac atcgagattg atacctttac
     1321 cggtatgagc agcagcaagg ataacccgaa ctacgatgac accaccatgc cgaaagagct
     1381 ggaagcgctg ctgcgttatt tcgaagcgag cgatagcgtg cagagcgcgc tgcgtgaggt
     1441 tatcgacttc caagaaaagt actgcaagta catcgagttc aagaccgaag acaagatgtt
     1501 caacgcgggt tttaacaaaa acctggcgtt tcagatcctg taccaaacct tcatgagccg
     1561 tagcttttgc cagacccaaa agggttatcg tgagatcggc ttccgtgaaa ttcaggatct
     1621 gtacgcgagc atgtactact tcatcaacct gggttaccaa gacttcgtga agaacctgct
     1681 gtttgagtgg accagcaacg tttacaaaat gggctatgcg aaccacaact tctactgggt
     1741 tggtaaacag ccgggcctgt atagcgatga cagcctgtgg ctgctgcaag cgtactatcg
     1801 ttacatcatt tataccaagg ataccagcat cctgaacgag gaagtgccgg ttgcggatgg
     1861 taacaacgaa aagcgtgcgg tgcgtgacac cctgaaagcg atcattcagt acagcgcgtt
     1921 catcagcgtt ggtaaccatg gtctgccgct gctggatctg gcggactgga acgattgcct
     1981 gaagattgat aacgactatg tggacggtgc gcagaaggag aaactgtact ataagcaact
     2041 gaaagaaacc ggtggcaaat acggcgatcg tttcatgagc gactatagcg agagcgttat
     2101 gaacgcgttt ctgctgaagg tggcggttga tgacctggcg gagatcgcgg aactggataa
     2161 ggacgcggag ctggaacaga gcatgcgtaa aatcagcagc gcgctgagcg aaaagattca
     2221 aaaatacgcg tggaagggta acttctttgc gcgtgtgctg ctgaaccgtt acaaagatgg
     2281 cagcttcacc tatgttggtg cgaagggcga caaactgagc gcggacccga acatcgatgg
     2341 tacctatttc ctgaacagct ttagctgggc gattctgagc ggtgtggcga ccgatgagca
     2401 gatcggcatt atggttgaca ccctgaagaa atacatgctg accccgtatg gcatccgtct
     2461 gaccaccccg gcggatctga acaaactggc gaacgacacc gcgaccgatc actacttcta
     2521 tggtgaccgt gaaaacggcg cggtgtttaa gcatgcgagc atgatggcga ccgcggcgct
     2581 ggcgaaggcg gcgaaaagcg ttaaggataa agcgctggcg aaagagatgg cgcgtctggc
     2641 gtactatatg atcgatctgg tgctgccgta caagaacctg gaagacccgt tcaaactggc
     2701 gggtaacccg cgtatctgca cccagtatat taacgcggac agcggcgaga acattggtcc
     2761 gctgctgagc ggtaccgcga cctggctgaa cctgaccatc accagcctgg cgggtattga
     2821 gtacgtgaag gatggcatta tgttcagccc ggttctgcgt gaggacgaaa cccgtctgga
     2881 atacagcctg aaggcgccgg gttgcagcta taacatcacc attaccaaac cgcgtggctt
     2941 tgcgcgtctg atcgacagcg cgtacgatct gcgtattgac ggtaagaaaa ccgacaactg
     3001 cattatcccg atgttcaccg atggtaaaga gcatagcgtg accctggagt tcaagtaaga
     3061 cgtcctagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgaagcttat
     3121 aatactcgag taaaaaaaat ccttagcttt cgctaaggat gtgcactcag aagaactcgt
     3181 caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg taaagcacga
     3241 ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta gccaacgcta
     3301 tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca gaaaagcggc
     3361 cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg agatcctcgc
     3421 cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc ccctgatgct
     3481 cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt gctcgctcga
     3541 tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc
     3601 gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat gacaggagat
     3661 cctgccccgg aacttcgccc aatagcagcc agtcccttcc cgcttcagtg acaacgtcga
     3721 gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct gcctcgtcct
     3781 ggagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg
     3841 ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc cagtcatagc
     3901 cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct tgttcaatca
     3961 tgtcgacttt ctcctcttta atagatcttc actgcccgct ttccagtcgg gaaacctgtc
     4021 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4081 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     4141 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     4201 gtttgatggt ggttaacggc gggatataac atgagctgtc ctcggtatcg tcgtatccca
     4261 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     4321 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     4381 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     4441 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     4501 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     4561 cgcccagtcg cgtaccgtcc tcatgggaga aaataatact gttgatgggt gtctggtcag
     4621 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     4681 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     4741 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     4801 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     4861 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     4921 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     4981 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     5041 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     5101 ggcgctatca tgccataccg cgaaaggttt tgcaccattc gatggtgtcg aattcctagc
     5161 ggagtgtata ctggcttact atgttggcac tgatgagggt gtcagtgaag tgcttcatgt
     5221 ggcaggagaa aaaaggctgc accggtgcgt cagcagaata tgtgatacag gatatattcc
     5281 gcttcctcgc tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct
     5341 tacgaacggg gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga
     5401 gggccgcggc aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct
     5461 gacgctcaaa tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5521 tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg tttaccggtg tcattccgct
     5581 gttatggccg cgtttgtctc attccacgcc tgacactcag ttccgggtag gcagttcgct
     5641 ccaagctgga ctgtatgcac gaaccccccg ttcagtccga ccgctgcgcc ttatccggta
     5701 actatcgtct tgagtccaac ccggaaagac atgcaaaagc accactggca gcagccactg
     5761 gtaattgatt tagaggagtt agtcttgaag tcatgcgccg gttaaggcta aactgaaagg
     5821 acaagttttg gtgactgcgc tcctccaagc cagttacctc ggttcaaaga gttggtagct
     5881 cagagaacct tcgaaaaacc gccctgcaag gcggtttttt cgttttcaga gcaagagatt
     5941 acgcgcagac caaaacgatc tcaagaagat catcttatta atcagataaa atatttctag
     6001 atttcagtgc aatttatctc ttcaaatgta gcacctgaag tcagccccat acgatataag
     6061 ttgtctgcag ttaatacgac tcactatagg gg
//
