LOCUS       sequence-193510-        7168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Tyrosine Ammonia Lyase from inducible T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7168)
  AUTHORS   Jones JA, Vernacchio VR, Collins SM, Shirke AN, Xiu Y, Englaender 
            JA, Cress BF, McCutcheon CC, Linhardt RJ, Gross RA, Koffas MAG
  TITLE     Complete Biosynthesis of Anthocyanins Using E. coli Polycultures.
  JOURNAL   MBio. 2017 Jun 6;8(3). pii: e00621-17. doi: 10.1128/mBio.00621-17.
  PUBMED    28588129
REFERENCE   2  (bases 1 to 7168)
  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: "MBio."; 
            date: "2017-06-6"; volume: "8(3). pii: e00621-17. doi"; pages: " 
            10.1128/mBio.00621-17"
COMMENT     Sequence Label: pETM6-RgTALsyn
FEATURES             Location/Qualifiers
     source          1..7168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     969..988
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1008..1085)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1291..1310
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1401..1420
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1401..1419
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1420..1444
                     /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             1459..1481
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3606..3624)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3620..3667
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3711..4166
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     4008..4029
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4281..5072,5073..5141))
                     /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     4904..4923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5142..5234)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5315..5903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5804..5823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6057..6074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6089..6228
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6154..6173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6314..6336
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6330..6521)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(7093..7168,1..1007))
                     /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"
ORIGIN
        1 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
       61 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      121 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
      181 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
      241 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
      301 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
      361 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
      421 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
      481 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
      541 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
      601 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
      661 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
      721 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
      781 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
      841 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
      901 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
      961 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1021 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     1081 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     1141 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     1201 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     1261 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     1321 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagccta ggatcgagat
     1381 cgatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     1441 ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat ggcgcctcgc
     1501 ccgacttcgc aaagccaggc ccgcacttgc ccgacgacgc aggttaccca agttgatatc
     1561 gttgagaaaa tgttggcggc tcctactgat agcacgctgg agctggacgg ttatagcctg
     1621 aatctgggtg atgtcgtgag cgctgcgcgt aagggtcgtc ctgtccgtgt caaagatagc
     1681 gatgaaatcc gcagcaaaat cgacaagagc gttgaattcc tgcgcagcca actgagcatg
     1741 tcggtttacg gtgtgacgac cggctttggc ggctccgcgg acacgcgcac ggaggacgca
     1801 attagcctgc aaaaggcgtt gctggaacac cagctgtgtg gtgtgttgcc gagcagcttc
     1861 gacagctttc gcttgggtcg tggtctggag aatagcctgc cgttggaagt cgttcgcggt
     1921 gcaatgacca ttcgtgtgaa ttcgctgacc cgtggccata gcgctgttcg tctggttgtt
     1981 ctggaagcac tgacgaactt tctgaaccac ggtattaccc cgattgttcc gctgcgcggt
     2041 acgatctccg cgagcggcga tctgtctcca ctgtcgtaca ttgcagcggc gattagcggt
     2101 cacccggata gcaaagttca cgtggtccat gaaggcaaag agaagatcct gtacgcgcgc
     2161 gaagcgatgg cgctgtttaa cctggagccg gtggttttgg gtccgaagga gggcctgggt
     2221 ctggtgaatg gtacggcagt ctccgcgagc atggcaacgc tggcactgca cgacgcgcac
     2281 atgttgagcc tgttgagcca atcgctgacc gcgatgaccg tggaggcgat ggtcggtcac
     2341 gcgggcagct tccatccatt cctgcacgat gttacgcgtc cgcacccgac gcaaatcgag
     2401 gtcgcgggta acattcgcaa actgctggag ggctcgcgct tcgcggtcca ccacgaggaa
     2461 gaggttaagg tcaaggatga tgaaggcatt ttgcgtcagg atcgttatcc gttgcgcacg
     2521 agcccgcaat ggttgggtcc gctggtgtcc gacctgattc acgctcatgc cgtcttgacg
     2581 atcgaagcgg gtcaaagcac caccgataac ccactgatcg atgttgagaa taagaccagc
     2641 catcacggtg gcaactttca agcggcagcg gttgccaaca cgatggaaaa gacccgtctg
     2701 ggcttggccc aaatcggtaa actgaatttc acccagctga cggagatgct gaacgcgggc
     2761 atgaatcgtg gcttgccgag ctgcctggcg gctgaagacc catccctgag ctatcattgc
     2821 aaaggtctgg acattgcggc ggctgcatat acgagcgaac tgggccacct ggctaacccg
     2881 gtcaccaccc acgtccaacc ggctgaaatg gcaaaccagg cggtgaatag cttggcgttg
     2941 attagcgcac gtcgtaccac ggaatctaac gacgttctgt ccctgctgct ggcaacgcac
     3001 ctgtactgcg tgctgcaggc gatcgacctg cgtgcgattg agttcgagtt caagaaacag
     3061 tttggtcctg ccattgttag cctgatcgac caacactttg gtagcgcgat gacgggtagc
     3121 aatctgcgtg atgagctggt tgaaaaggtc aataagactc tggccaagcg tttggagcaa
     3181 accaatagct acgatctggt tccgcgctgg cacgacgctt ttagcttcgc tgcaggcact
     3241 gttgtcgagg ttctgtccag cacgagcctg agcttggcgg ccgtgaacgc atggaaggtt
     3301 gcggcagccg agagcgcgat ctccttgacg cgccaggtcc gtgaaacgtt ttggtccgct
     3361 gcaagcacct ccagcccggc gttgtcttac ttgagcccgc gcacgcagat cctgtacgca
     3421 tttgtgcgtg aggaactggg tgtcaaagcc cgccgtggtg acgtcttctt gggtaaacaa
     3481 gaagttacca tcggcagcaa cgttagcaag atttacgaag ccatcaagag cggccgtatc
     3541 aacaatgttc tgctgaagat gctggcataa actagtcgca gcttaattaa cctaaactgc
     3601 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3661 ttttttgcta gcgaaaggag gagtcgacta tatccggatt ggcgaatggg acgcgccctg
     3721 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3781 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3841 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3901 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3961 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4021 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4081 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4141 taacaaaata ttaacgttta caatttctgg cggcacgatg gcatgagatt atcaaaaagg
     4201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatca
     5161 tgattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5221 gaaaaataaa caaataggtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     5281 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     5341 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5401 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5461 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5521 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5581 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5641 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5701 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5761 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5821 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5881 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5941 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     6001 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     6061 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acacatctgt
     6121 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6181 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     6241 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     6301 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     6361 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     6421 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     6481 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     6541 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     6601 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     6661 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     6721 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     6781 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     6841 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     6901 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     6961 gacaggagca cgatcatgct agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     7021 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     7081 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7141 tgaatcggcc aacgcgcggg gagaggcg
//
