LOCUS       sequence-77277-e        9393 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Origin Of Replication: p15a, Selection Markers: Amp, Promoters: 
            PLacUV5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9393)
  AUTHORS   Juminaga D, Baidoo EE, Redding-Johanson AM, Batth TS, Burd H, 
            Mukhopadhyay A, Petzold CJ, Keasling JD
  TITLE     Modular engineering of L-tyrosine production in Escherichia coli.
  JOURNAL   Appl Environ Microbiol. 2012 Jan;78(1):89-98. doi: 
            10.1128/AEM.06017-11. Epub 2011 Oct 21.
  PUBMED    22020510
REFERENCE   2  (bases 1 to 9393)
  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"; doi: 
            "10.1128/AEM.06017-11"; journalName: "Appl Environ Microbiol"; date:
            "2012-01"; volume: "78"; issue: "1"; pages: "89-98"
COMMENT     Sequence Label: pY3
FEATURES             Location/Qualifiers
     source          1..9393
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(49..1131)
                     /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     1093..1112
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1132..1209)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1415..1434
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(1520..1525,1526..1543,1544..1550)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1558..1574
                     /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)."
     terminator      5077..5148
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5164..5191
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        join(5404..5409,5410..5426,5427..5433)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5441..5457
                     /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)."
     terminator      7344..7415
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7431..7458
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(7486..7503)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7620..8165)
                     /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(7700..7719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(8279..8373)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(8403..9194,9195..9263))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     9026..9045
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9264..9368)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
       61 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      121 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
      181 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
      241 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
      301 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
      361 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
      421 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
      481 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
      541 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
      601 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
      661 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
      721 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
      781 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
      841 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
      901 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
      961 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     1021 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     1081 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     1141 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     1201 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     1261 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     1321 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     1381 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     1441 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     1501 tcgtttaggc accccaggct ttacacttta tgcttccggc tcgtataatg tgtggaattg
     1561 tgagcggata acaatttcag aattcaaaag atctaaagga ggccatccgt gtttcaaaaa
     1621 gttgacgcct acgctggcga cccgattctt acgcttatgg agcgttttaa agaagaccct
     1681 cgcagcgaca aagtgaattt aagtatcggt ctgtactaca acgaagacgg aattattcca
     1741 caactgcaag ccgtggcgga ggcggaagcg cgcctgaatg cgcagcctca tggcgcttcg
     1801 ctttatttac cgatggaagg gcttaactgc tatcgccatg ccattgcgcc gctgctgttt
     1861 ggtgcggacc atccggtact gaaacaacag cgcgtagcaa ccattcaaac ccttggcggc
     1921 tccggggcat tgaaagtggg cgcggatttc ctgaaacgct acttcccgga atcaggcgtc
     1981 tgggtcagcg atcctacctg ggaaaaccac gtagcaatat tcgccggggc tggattcgaa
     2041 gtgagtactt acccctggta tgacgaagcg actaacggcg tgcgctttaa tgacctgttg
     2101 gcgacgctga aaacattacc tgcccgcagt attgtgttgc tgcatccatg ttgccacaac
     2161 ccaacgggtg ccgatctcac taatgatcag tgggatgcgg tgattgaaat tctcaaagcc
     2221 cgcgagctta ttccattcct cgatattgcc tatcaaggat ttggtgccgg tatggaagag
     2281 gatgcctacg ctattcgcgc cattgccagc gctggattac ccgctctggt gagcaattcg
     2341 ttctcgaaaa ttttctccct ttacggcgag cgcgtcggcg gactttctgt tatgtgtgaa
     2401 gatgccgaag ccgctggccg cgtactgggg caattgaaag caacagttcg ccgcaactac
     2461 tccagcccgc cgaattttgg tgcgcaggtg gtggctgcag tgctgaatga cgaggcattg
     2521 aaagccagct ggctggcgga agtagaagag atgcgtactc gcattctggc aatgcgtcag
     2581 gaattggtga aggtattaag cacagagatg ccagaacgca atttcgatta tctgcttaat
     2641 cagcgcggca tgttcagtta taccggttta agtgccgctc aggttgaccg actacgtgaa
     2701 gaatttggtg tctatctcat cgccagcggt cgcatgtgtg tcgccgggtt aaatacggca
     2761 aatgtacaac gtgtggcaaa ggcgtttgct gcggtgatgt aaggatctaa aggaggccat
     2821 ccatggttgc tgaattgacc gcattacgcg atcaaattga tgaagtcgat aaagcgctgc
     2881 tgaatttatt agcgaagcgt ctggaactgg ttgctgaagt gggcgaggtg aaaagccgct
     2941 ttggactgcc tatttatgtt ccggagcgcg aggcatctat gttggcctcg cgtcgtgcag
     3001 aggcggaagc tctgggtgta ccgccggatc tgattgagga tgttttgcgt cgggtgatgc
     3061 gtgaatctta ctccagtgaa aacgacaaag gatttaaaac actttgtccg tcactgcgtc
     3121 cggtggttat cgtcggcggt ggcggtcaga tgggacgcct gttcgagaag atgctgaccc
     3181 tctcgggtta tcaggtgcgg attctggagc aacatgactg ggatcgagcg gctgatattg
     3241 ttgccgatgc cggaatggtg attgttagtg tgccaatcca cgttactgag caagttattg
     3301 gcaaattacc gcctttaccg aaagattgta ttctggtcga tctggcatca gtgaaaaatg
     3361 ggccattaca ggccatgctg gtggcgcatg atggtccggt gctggggcta cacccgatgt
     3421 tcggtccgga cagcggtagc ctggcaaagc aagttgtggt ctggtgtgat ggacgtaaac
     3481 cggaagcata ccaatggttt ctggagcaaa ttcaggtctg gggcgctcgg ctgcatcgta
     3541 ttagcgccgt cgagcacgat cagaatatgg cgtttattca ggcactgcgc cactttgcta
     3601 cttttgctta cgggctgcac ctggcagaag aaaatgttca gcttgagcaa cttctggcgc
     3661 tctcttcgcc gatttaccgc cttgagctgg cgatggtcgg gcgactgttt gctcaggacc
     3721 cgcagcttta tgccgacatc attatgtcgt cagagcgtaa tctggcgtta atcaaacgtt
     3781 actataagcg tttcggcgag gcgattgagt tgctggagca gggcgataag caggcgttta
     3841 ttgacagttt ccgcaaggtg gagcactggt tcggcgatta cgcacagcgt tttcagagtg
     3901 aaagccgcgt gttattgcgt caggcgaatg acaatcgcca gtaaggatct aaaggaggcc
     3961 atccatggct ggaaacacaa ttggacaact ctttcgcgta accaccttcg gcgaatcgca
     4021 cgggctggcg ctcggctgca tcgtcgatgg tgttccgcca ggcattccgc tgacggaagc
     4081 ggacctgcaa catgacctcg accgtcgtcg ccctgggaca tcgcgctata ccacccagcg
     4141 ccgcgagccg gatcaggtca aaattctctc cggtgttttt gaaggcgtta ctaccggcac
     4201 cagcattggc ttgttgatcg aaaacactga ccagcgctct caggattaca gtgcgattaa
     4261 ggacgttttc cgtccaggcc atgccgatta cacctacgaa caaaaatacg gtctgcgcga
     4321 ttatcgcggc ggtggacgtt cttccgcccg cgaaaccgcc atgcgcgtgg cggcaggagc
     4381 tattgccaaa aaatatctcg ccgagaaatt tggtattgaa atccgtggct gcctgaccca
     4441 gatgggcgac attccgctgg atatcaaaga ctggtcgcag gtcgagcaaa atccgttttt
     4501 ttgcccggac cccgacaaaa tcgacgcgtt agacgagttg atgcgtgcgc tgaaaaaaga
     4561 gggcgactcc atcggcgcta aagtcaccgt tgttgccagt ggcgttcctg ccggacttgg
     4621 cgagccggtc tttgaccgcc tggatgctga catcgcccat gcgctgatga gcatcaacgc
     4681 ggtgaaaggc gtggaaattg gcgacggctt tgacgtggtg gcgctgcgcg gcagccagaa
     4741 ccgcgatgaa atcaccaaag acggtttcca gagcaaccat gcgggcggca ttctcggcgg
     4801 tatcagcagc gggcagcaaa tcattgccca tatggcgctg aaaccgacct ccagcattac
     4861 cgtgccgggt cgtaccatta accgctttgg cgaagaagtt gagatgatca ccaaaggccg
     4921 tcacgatccc tgtgtcggta tccgcgcagt gccgatcgca gaagcgatgc tggcgatcgt
     4981 tttaatggat cacctgttac ggcaacgggc gcaaaatgcc gatgtgaaga ctgatattcc
     5041 acgctggtaa ggatctaatt aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg
     5101 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
     5161 actggctcac cttcgggtgg gcctttctgc gtttatagcg aattggatct ggtttgacag
     5221 cttatcatcg actgcacggt gcaccaatgc ttctggcgtc aggcagccat cggaagctgt
     5281 ggtatggctg tgcaggtcgt aaatcactgc ataattcgtg tcgctcaagg cgcactcccg
     5341 ttctggataa tgttttttgc gccgacatca taacggttct ggcaaatatt ctgaaatgag
     5401 ctgttgacaa ttaatcatcc ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt
     5461 caggatctaa aggaggccat ccatggaatc cctgacgtta caacccatcg ctcgtgtcga
     5521 tggcactatt aatctgcccg gttccaagag cgtttctaac cgcgctttat tgctggcggc
     5581 attagcacac ggcaaaacag tattaaccaa tctgctggat agcgatgacg tgcgccatat
     5641 gctgaatgca ttaacagcgt taggggtaag ctatacgctt tcagccgatc gtacgcgttg
     5701 cgaaattatc ggtaacggcg gtccattaca cgcagaaggt gccctggagt tgttcctcgg
     5761 taacgccgga acggcaatgc gtccgctggc ggcagctctt tgtctgggta gcaatgatat
     5821 tgtgctgacc ggtgagccgc gtatgaaaga acgcccgatt ggtcatctgg tggatgcgct
     5881 gcgcctgggc ggggcgaaga tcacttacct ggaacaagaa aattatccgc cgttgcgttt
     5941 acagggcggc tttactggcg gcaacgttga cgttgatggc tccgtttcca gccaattcct
     6001 caccgcactg ttaatgactg cgcctcttgc gccggaagat acggtgattc gtattaaagg
     6061 cgatctggtt tctaaacctt atatcgacat cacactcaat ctgatgaaga cgtttggtgt
     6121 tgaaattgaa aatcagcact atcaacaatt tgtcgtaaaa ggcgggcagt cttatcagtc
     6181 tccgggtact tatttggtcg aaggcgatgc atcttcggct tcttactttc tggcagcagc
     6241 agcaatcaaa ggcggcactg taaaagtgac cggtattgga cgtaacagta tgcagggtga
     6301 tattcgcttt gctgatgtgc tggaaaaaat gggcgcgacc atttgctggg gcgatgatta
     6361 tatttcctgc acgcgtggtg aactgaacgc tattgatatg gatatgaacc atattcctga
     6421 tgcggcgatg accattgcca cggcggcgtt atttgcaaaa ggcaccacca cgctgcgcaa
     6481 tatctataac tggcgtgtta aagagaccga tcgcctgttt gcgatggcaa cagaactgcg
     6541 taaagtcggc gcggaagtgg aagaggggca cgattacatt cgtatcactc ctccggaaaa
     6601 actgaacttt gccgagatcg cgacatacaa tgatcaccgg atggcgatgt gtttctcgct
     6661 ggtggcgttg tcagatacac cagtgacgat tcttgatccc aaatgcacgg ccaaaacatt
     6721 tccggattat ttcgagcagc tggcgcggat tagccaggca gcctgaggat ctaaaggagg
     6781 ccatccatga cacaacctct ttttctgatc gggcctcggg gctgtggtaa aacaacggtc
     6841 ggaatggccc ttgccgattc gcttaaccgt cggtttgtcg ataccgatca gtggttgcaa
     6901 tcacagctca atatgacggt cgcggagatc gtcgaaaggg aagagtgggc gggatttcgc
     6961 gccagagaaa cggcggcgct ggaagcggta actgcgccat ccaccgttat cgctacaggc
     7021 ggcggcatta ttctgacgga atttaatcgt cacttcatgc aaaataacgg gatcgtggtt
     7081 tatttgtgtg cgccagtatc agtcctggtt aaccgactgc aagctgcacc ggaagaagat
     7141 ttacggccaa ccttaacggg aaaaccgctg agcgaagaag ttcaggaagt gctggaagaa
     7201 cgcgatgcgc tatatcgcga agttgcgcat attatcatcg acgcaacaaa cgaacccagc
     7261 caggtgattt ctgaaattcg cagcgccctg gcacagacga tcaattgttg aggatccaaa
     7321 ctcgagtaag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     7381 ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt
     7441 cgggtgggcc tttctgcgtt tatacctagg gatatattcc gcttcctcgc tcactgactc
     7501 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
     7561 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     7621 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     7681 cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc
     7741 tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct
     7801 cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca
     7861 cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     7921 cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt
     7981 tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg
     8041 ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac
     8101 cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat
     8161 ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct
     8221 cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgttacta gtgcttggat
     8281 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     8341 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgtaaac ttggtctgac
     8401 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     8461 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     8521 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     8581 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     8641 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     8701 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     8761 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     8821 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     8881 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     8941 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     9001 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     9061 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     9121 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     9181 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     9241 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     9301 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     9361 gttccgcgca catttccccg aaaagtgcca cct
//
