LOCUS       sequence-384049-        7814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible AIDA-Tyrosinase expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7814)
  AUTHORS   Binelli MR, Kan A, Rozas LEA, Pisaturo G, Prakash N, Studart AR
  TITLE     Complex Living Materials made by Light-based Printing of Genetically
            Programmed Bacteria.
  JOURNAL   Adv Mater. 2022 Nov 29:e2207483. doi: 10.1002/adma.202207483.
  PUBMED    36444840
REFERENCE   2  (bases 1 to 7814)
  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: "Adv Mater. 
            2022 Nov 29:e2207483. doi: 10.1002/adma.202207483."
COMMENT     Sequence Label: pLN-AIDA-Tyr1
FEATURES             Location/Qualifiers
     source          1..7814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             191..1273
                     /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(210..229)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     terminator      1293..1364
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1380..1407
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    1428..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)."
     protein_bind    1451..1467
                     /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)."
     misc_RNA        1483..1532
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             1735..1752
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2677..2697
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2698..2727
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     terminator      complement(4326..4368)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     terminator      4475..4506
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4605..5420)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5328..5347
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5412..5431
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(5778..6728)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      6776..6998
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(7490..7584)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      complement(7740..7769)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 cattactcgc atccattctc aggctgtctc gtctcgtctc gctgggagtt cgtagacgga
       61 aacaaacgca gaatccaagc cggaattcgg acccacaagg cggcgcgcca tcgaatggcg
      121 caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcgcttca aggtggtgca
      181 tatggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
      241 tcagaccgtt tctcgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
      301 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
      361 gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc acgcgccgtc
      421 gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc
      481 gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca
      541 acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca ttgctgtgga
      601 agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga cacccatcaa
      661 cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc tggtcgcatt
      721 gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg
      781 tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga
      841 aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat
      901 cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat
      961 taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg acgataccga
     1021 agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg
     1081 gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca
     1141 gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc
     1201 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     1261 aagcgggcag taataatact agagccaggc atcaaataaa acgaaaggct cagtcgaaag
     1321 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     1381 gctcaccttc gggtgggcct ttctgcgttt atatactagt agcggaattg tgagcggata
     1441 acaattgaca ttgtgagcgg ataacaagat actgagcaca agctgtcacc ggatgtgctt
     1501 tccggtctga tgagtccgtg aggacgaaac agcctctaca aataattttg tttaatggta
     1561 gcgacagccg agaaatactc gatgaataag gcctacagta tcatttggag ccactccaga
     1621 caggcctgga ttgtggcctc agagttagcc agaggacatg gttttgtcct tgcaaaaaat
     1681 acactgctgg tattggcggt tgtttccaca atcggaaatg catttgcagt cgaccaccat
     1741 caccatcacc atctggaagc gctgttccag ggtccgggta ccggtaataa atatcgcgtt
     1801 cgtaaaaatg tgttgcatct gaccgacacc gaaaaacgcg atttcgtgcg taccgttctg
     1861 attctcaaag agaaaggtat ctacgaccgc tatatcgcct ggcatggcgc agcgggtaaa
     1921 tttcatactc ctcctggcag cgatcgcaac gccgcgcata tgagcagcgc cttcctgccg
     1981 tggcaccgcg aatatctgct gcgttttgaa cgcgatctgc aaagcatcaa cccagaagtg
     2041 accctgccgt actgggagtg ggaaaccgat gcgcaaatgc aggatccgag tcagtcgcag
     2101 atctggtcgg cggactttat gggcggcaac ggtaacccga tcaaagattt tattgtcgat
     2161 accggtccgt tcgccgcagg ccgctggacc accattgatg aacagggcaa tccgagcggc
     2221 ggcctgaagc gcaattttgg cgccaccaaa gaagcgccga cgctgccgac ccgagatgac
     2281 gtgctgaacg cgctgaaaat tacccagtat gacacgccgc cgtgggatat gaccagccag
     2341 aatagctttc gtaaccaact ggaaggtttt atcaatggcc cgcagctgca caaccgcgtg
     2401 catcgttggg ttggtggtca aatgggcgta gtcccaactg cgccgaacga tccagtattt
     2461 ttcctgcatc acgccaacgt ggatcgaatc tgggcggtct ggcagattat tcacaggaat
     2521 cagaattatc agccaatgaa aaacggtccg ttcggtcaga acttccgcga cccgatgtat
     2581 ccgtggaaca ccaccccaga agatgttatg aaccatcgca aactgggcta tgtctacgat
     2641 atcgaactgc gcaaaagcaa acgttccagc gagctcgaaa acctgtactt ccagggtgaa
     2701 cagaaactga ttagcgaaga agatctgtct agagtgaata acaatggaag cattgtcatt
     2761 aataacagca ttataaacgg gaatattacg aatgatgctg acttaagttt tggtacagca
     2821 aagctgctct ctgctacagt gaatggtagt cttgttaata acaaaaatat cattcttaat
     2881 cctacaaaag aaagtgcggc cgctataggt aatactctta ccgtgtcaaa ttatactggg
     2941 acaccgggaa gtgttatttc tcttggtggt gtgcttgaag gagataattc acttacggac
     3001 cgtctggtgg tgaaaggtaa tacctctggt caaagtgaca tcgtttatgt caatgaagat
     3061 ggcagtggtg gtcagacgag agatggtatt aatattattt ctgtagaggg aaattctgat
     3121 gcagaattct ctctgaagaa ccgcgtagtt gccggagctt atgattacac actgcagaaa
     3181 ggaaacgaga gtgggacaga taataaggga tggtatttaa ccagtcatct tcccacatct
     3241 gatacccggc aatacagacc ggagaacgga agttatgcta ccaatatggc actggctaac
     3301 tcactgttcc tcatggattt gaatgagcgt aagcaattca gggccatgag tgataataca
     3361 cagcctgagt ctgcatccgt gtggatgaag atcactggag gaataagctc tggtaagctg
     3421 aatgacgggc aaaataaaac aacaaccaat cagtttatca atcagctcgg gggggatatt
     3481 tataaattcc atgctgaaca actgggtgat tttaccttag ggattatggg aggatacgcg
     3541 aatgcaaaag gtaaaacgat aaattacacg agcaacaaag ctgccagaaa cacactggat
     3601 ggttattctg tcggggtata cggtacgtgg tatcagaatg gggaaaatgc aacagggctc
     3661 tttgctgaaa cttggatgca atataactgg tttaatgcat cagtgaaagg tgacggactg
     3721 gaagaagaaa aatataatct gaatggttta accgcttctg caggtggggg atataacctg
     3781 aatgtgcaca catggacatc acctgaagga ataacaggtg aattctggtt acagcctcat
     3841 ttgcaggctg tctggatggg ggttacaccg gatacacatc aggaggataa cggaacggtg
     3901 gtgcagggag cagggaaaaa taatattcag acaaaagcag gtattcgtgc atcctggaag
     3961 gtgaaaagca ccctggataa ggataccggg cggaggttcc gtccgtatat agaggcaaac
     4021 tggatccata acactcatga atttggtgtt aaaatgagtg atgacagcca gttgttgtca
     4081 ggtagccgaa atcagggaga gataaagaca ggtattgaag gggtgattac tcaaaacttg
     4141 tcagtgaatg gcggagtcgc atatcaggca ggaggtcacg ggagcaatgc catctccgga
     4201 gcactgggga taaaatacag cttctgataa ctcggtacca aattccagaa aagaggcctc
     4261 ccgaaagggg ggcctttttt cgttttggtc caaaaaagcc cgcacctgac agtgcgggct
     4321 ttttttttca gataaaaaaa atccttagct ttcgctaagg atgatttctg cactgaaggt
     4381 cctcaatcgc actggaaaca tcaaggtcgc caggatacat agattaccac aactccgagc
     4441 ccttccaccc gccggcgaga cctgattagc agaaagtcaa aagcctccga ccggaggctt
     4501 ttgactaaaa cttcccttgg ggttatcatt ggggctcact caaaggcggt aatcagataa
     4561 aaaaaatcct tagctttcgc taaggatgat ttctgctagt attattagaa aaactcatcg
     4621 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     4681 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccaaagaat ggcaaggtcc
     4741 tggtaacggt ctgcgattcc gacccgtcca acatcaatac aacctattaa tttcccctcg
     4801 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     4861 ggcaagagct tgtgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     4921 tcaaaatcac tcgcatcaac caaaccgtta ttcatgcgtg attgcgcctg agcaagacga
     4981 aatacacgat cgctgttaaa aggacaatta caaacaggaa tcgaatgtaa ccggcgcagg
     5041 aacacggcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     5101 aaggctgttt tcccaggaat cgcggtggtg agtaaccacg catcatcagg agtacggata
     5161 aaatgcttga tggtcgggag aggcataaac tccgtcagcc agttgagacg gaccatctca
     5221 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     5281 ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     5341 ttatacccat ataaatcagc gtccatgttg gagtttaagc gcggacggga gcaagacgtt
     5401 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     5461 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     5521 acgtggcttt gttgaataaa tcgaactttt gctgagttga aggatcagct ctagtagtta
     5581 cattgtcgat ctgttcatgg tgaacagctt tgaatgcacc aaaaactcgt aaaagctctg
     5641 atgtatctat cttttttaca ccgttttcat ctgtgcatat ggacagtttt ccctttgata
     5701 tgtaacggtg aacagttgtt ctacttttgt ttgttagtct tgatgcttca ctgatagata
     5761 caagagccat aagaacctca gatccttccg tatttagcca gtatgttctc tagtgtggtt
     5821 cgttgttttt gcgtgagcca tgagaacgaa ccattgagat catacttact ttgcatgtca
     5881 ctcaaaaatt ttgcctcaaa actggtgagc tgaatttttg cagttaaagc atcgtgtagt
     5941 gtttttctta gtccgttatg taggtaggaa tctgatgtaa tggttgttgg tattttgtca
     6001 ccattcattt ttatctggtt gttctcaagt tcggttacga gatccatttg tctatctagt
     6061 tcaacttgga aaatcaacgt atcagtcggg cggcctcgct tatcaaccac caatttcata
     6121 ttgctgtaag tgtttaaatc tttacttatt ggtttcaaaa cccattggtt aagcctttta
     6181 aactcatggt agttattttc aagcattaac atgaacttaa attcatcaag gctaatctct
     6241 atatttgcct tgtgagtttt cttttgtgtt agttctttta ataaccactc ataaatcctc
     6301 atagagtatt tgttttcaaa agacttaaca tgttccagat tatattttat gaattttttt
     6361 aactggaaaa gataaggcaa tatctcttca ctaaaaacta attctaattt ttcgcttgag
     6421 aacttggcat agtttgtcca ctggaaaatc ccaaagcctt taaccaaagg attcctgatt
     6481 tccacagttc tcgtcatcag ctctctggtt gctttagcta atacaccata agcattttcc
     6541 ctactgatgt tcatcatctg agcgtattgg ttataagtga acgataccgt ccgttctttc
     6601 cttgtagggt tttcaatcgt ggggttgagt agtgccacac agcataaaat tagcttggtt
     6661 tcatgctccg ttaagtcata gcgactaatc gctagttcat ttgctttgaa aacaactaat
     6721 tcagacatac atctcaattg gtctaggtga ttttaatcac tataccaatt gagatgggct
     6781 agtcaatgat aattactagt ccttttcccg ggagatctgg gtatctgtaa attctgctag
     6841 acctttgctg gaaaacttgt aaattctgct agaccctctg taaattccgc tagacctttg
     6901 tgtgtttttt ttgtttatat tcaagtggtt ataatttata gaataaagaa agaataaaaa
     6961 aagataaaaa gaatagatcc cagccctgtg tataactcac tactttagtc agttccccag
     7021 tattacaaaa ggatgtcgca aacgctgttt gctcctctac aaaacagacc ttaaaaccct
     7081 aaaggcttaa gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc
     7141 gaccatcagg cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc
     7201 tctggcagtg aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa
     7261 actacccata atacaagaaa agcccgtcac gggcttctca tggcgtttta tggcgggtct
     7321 gctatgtggt gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt
     7381 ctgaccactt cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg
     7441 cagcggtatc atcaacaggc ttacccgtct tactgtccct agtgcttgga ttctcaccaa
     7501 taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca
     7561 ttactggatc tatcaacagg agtccaagcg agctcgtaaa cttggtctga cagctctagc
     7621 tccggcaaaa aaacgggcaa ggtgtcacca ccctgccctt tttctttaaa accgaaaaga
     7681 ttacttcgcg tttgccacct gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc
     7741 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgctacgtaa ttagttagtt
     7801 agcccttagt gacg
//
