LOCUS       sequence-142936-        6666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of ectABC in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6666)
  AUTHORS   Ning Y, Wu X, Zhang C, Xu Q, Chen N, Xie X
  TITLE     Pathway construction and metabolic engineering for fermentative 
            production of ectoine in Escherichia coli.
  JOURNAL   Metab Eng. 2016 Mar 9;36:10-18. doi: 10.1016/j.ymben.2016.02.013.
  PUBMED    26969253
REFERENCE   2  (bases 1 to 6666)
  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: "Metab 
            Eng."; date: "2016-03-9"; volume: "36:10-18. doi"; pages: " 
            10.1016/j.ymben.2016.02.013"
COMMENT     Sequence Label: pTrcECT
FEATURES             Location/Qualifiers
     source          1..6666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(193..198,199..215,216..222)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    230..246
                     /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)."
     primer_bind     235..257
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     complement(2849..2866)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3019..3105
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3197..3224
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3244..3335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3336..3404,3405..4196)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3554..3573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4367..4955
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4856..4875
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5109..5126
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5141..5281
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5207..5226)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5367..5389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5467..5544
                     /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."
     CDS             5545..6627
                     /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     complement(5564..5583)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gctcgtataa tgtgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgg aattcatgaa cgcaaccaca gagcccttta
      301 caccctccgc cgacctggcc aagcccagcg tggccgatgc cgtggtcggc catgaggcct
      361 caccgctctt catccgcaag ccaagccccg atgacggctg gggcatctac gagctggtca
      421 agtcctgtcc gcctctcgac gtcaattccg cctacgccta tctgttgctg gccacccagt
      481 tccgcgatag ctgcgccgtg gcgaccaacg aagagggcga gatcgtcggc ttcgtttccg
      541 gctacgtgaa gagcaacgcc cccgatacct atttcctctg gcaggttgcc gtgggcgaga
      601 aggcacgtgg caccggcctg gcccgtcgtc tggtggaagc cgtgatgaca cgcccggaaa
      661 tggccgaggt ccaccatctc gagaccacta tcacgcccga caaccaggcg tcctggggct
      721 tgttccgccg tctcgccgat cgctggcagg cgccgttgaa cagccgcgaa tacttctcca
      781 ccgatcaact cggcggtgag catgacccgg aaaacctcgt tcgcatcggc ccgttccaga
      841 ccgaccagat ctgagccggg acgccgcctg gccggcccgg tacgggccgg caacccgtct
      901 tttcgtttta tcactttccc ccacaggagg tcgcaatgca gacccagatt ctcgaacgca
      961 tggagtccga cgttcggacc tactcccgct ccttcccggt cgtcttcacc aaggcgcgca
     1021 atgcccgcct gaccgacgag gaagggcgcg agtacatcga cttcctggcc ggtgccggca
     1081 ccctgaacta cggccacaac aacccgcacc tcaagcaggc gctgctcgac tatatcgaca
     1141 gcgacggcat cgtccacggc ctggacttct ggactgcggc caagcgcgac tatctggaaa
     1201 ccctggaaga ggtgatcctc aagccgcgcg gtctcgacta caaggtgcat ctgcccggac
     1261 cgactggcac caacgccgtc gaggcggcca ttcgcctggc ccgggtcgcc aaggggcgcc
     1321 acaatatcgt ctccttcacc aacggctttc atggcgtcac catgggcgcg ctggcgacca
     1381 ccggtaaccg caagttccgc gaggccaccg gtggcgtgcc gacccaggct gcttccttca
     1441 tgccgttcga tggctacctc ggcagcagca ccgacaccct cgactacttc gagaagctgc
     1501 tcggcgacaa gtccggcggc ctggacgtgc ccgcggcggt gatcgtcgag acagtgcagg
     1561 gcgagggcgg tatcaatgtc gccggcctgg agtggctcaa gcgcctcgag agcatctgcc
     1621 gcgccaatga catcctgctg atcatcgacg acatccaggc gggctgcggc cggaccggca
     1681 agttcttcag cttcgagcat gccggcatca cgccggatat cgtgaccaac tccaagtcgc
     1741 tgtccggtta cggcctgccg ttcgctcacg tcctgatgcg ccccgagctc gacaagtgga
     1801 agcccggtca gtacaacggc accttccgcg gcttcaacct ggctttcgcc actgctgctg
     1861 ccgccatgcg caagtactgg agcgacgaca ccttcgagcg tgacgtgcag cgcaaggctc
     1921 gcatcgtcga ggaacgcttc ggcaagatcg ccgcctggct gagcgagaac ggcatcgagg
     1981 cctccgagcg cggccgcggg ctgatgcggg gcatcgacgt gggttccggc gatatcgccg
     2041 acaagatcac ccaccaagcc ttcgagaacg ggttgatcat cgaaaccagc ggtcaggacg
     2101 gcgaagtggt caagtgcctg tgcccgctga ccattcccga cgaagacctg gtcgagggac
     2161 tcgacatcct cgagaccagc accaagcagg cctttagctg atcgcctgag gtgcgccatc
     2221 gggcctgtcc atggcatcct gtatcggtcg gccgtgcgcg gccggccagt cattgattca
     2281 ctggagaatc gacatgatcg ttcgcaatct cgaagaagcg cgccagaccg accgtctggt
     2341 caccgccgaa aacggcaact gggacagcac ccgcctgtcg ctggccgaag atggtggcaa
     2401 ctgctccttc cacatcaccc gcatcttcga gggtaccgag acccacatcc actataagca
     2461 tcacttcgag gctgtttatt gcatcgaagg cgagggcgaa gtggaaaccc tggccgatgg
     2521 caagatctgg cccatcaagc cgggtgacat ctacatcctc gaccagcacg acgagcacct
     2581 gctgcgcgcc agcaagacca tgcacctggc ctgcgtgttc acgccgggcc tgaccggcaa
     2641 cgaagtgcac cgcgaagacg gttcctacgc acctgccgac gaagccgacg accagaagcc
     2701 gctgtaaccc ggcgcagtat tctgccgtct cgcacgaaga gcccccggtc acgatcgggg
     2761 gctctttcgt tgttcgcagc ggatcctcta gagtcgacct gcaggcatgc aagcttggct
     2821 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
     2881 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     2941 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     3001 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     3061 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     3121 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     3181 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     3241 ctttttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3541 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     3601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3721 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3841 aagccatacc aaacgacgag cgtgacacca cgatgcctac agcaatggca acaacgttgc
     3901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4201 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4261 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4321 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4381 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4441 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4501 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4561 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4621 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4681 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4741 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4801 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4861 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4921 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4981 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5041 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5101 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     5161 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     5221 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     5281 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     5341 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     5401 atcaccgaaa cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcatt
     5461 tacgttgaca ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     5521 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     5581 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     5641 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     5701 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     5761 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     5821 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     5881 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     5941 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     6001 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     6061 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     6121 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     6181 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     6241 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     6301 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     6361 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     6421 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     6481 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     6541 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     6601 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agcgcgaatt
     6661 gatctg
//
