LOCUS       sequence-95205-e        4430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PTac alpha fragment expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4430)
  AUTHORS   Segall-Shapiro TH, Meyer AJ, Ellington AD, Sontag ED, Voigt CA
  TITLE     A 'resource allocator' for transcription based on a highly 
            fragmented T7 RNA polymerase.
  JOURNAL   Mol Syst Biol. 2014 Jul 30;10(7):742.
  PUBMED    25080493
REFERENCE   2  (bases 1 to 4430)
  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: "Mol Syst 
            Biol."; date: "2014-07-30"; volume: "10(7)"; pages: "742"
COMMENT     Sequence Label: pTHSSd_29
FEATURES             Location/Qualifiers
     source          1..4430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      248..279
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     rep_origin      790..1334
                     /direction=RIGHT
                     /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     1235..1254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1451..1468
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             2043..2834
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      complement(2961..2990)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     CDS             complement(3053..4135)
                     /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     4097..4116
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4270..4275,4276..4291,4292..4298)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    4304..4323
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     misc_RNA        4339..4388
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
ORIGIN
        1 atgaacacga ttaacatcgc taagaacgac ttctctgaca tcgaactggc tgctatcccg
       61 ttcaacactc tggctgacca ttacggtgag cgtttagctc gcgaacagtt ggcccttgag
      121 catgagtctt acgagatggg tgaagcacgc ttccgcaaga tgtttgagcg tcaacttaaa
      181 gctggtgagg ttgcggataa ctaacgctgc aggagtcact aagggttagt tagttagatt
      241 agcagaaagt caaaagcctc cgaccggagg cttttgacta aaacttccct tggggttatc
      301 attggggctc actcaaaggc ggtaatcaga taaaaaaaat ccttagcttt cgctaaggat
      361 gatttctgct agagatggaa tagactggat ggaggcggat aaagttgcag gaccacttct
      421 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      481 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      541 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      601 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
      661 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
      721 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccttaataag
      781 atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac
      841 cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa
      901 ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca
      961 tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg
     1021 catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact
     1081 gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg
     1141 tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag
     1201 gcaggaacag gagagcgcac gagggagccg ccaggggaaa cgcctggtat ctttatagtc
     1261 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     1321 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     1381 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     1441 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg
     1501 ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca
     1561 tcagtgccaa catagtaagc cagtatacac tccgctagcg ctgaggtctg cctcgtgaag
     1621 aaggtgttgc tgactcatac caggcctgaa tcgccccatc atccagccag aaagtgaggg
     1681 agccacggtt gatgagagct ttgttgtagg tggaccagtt ggtgattttg aacttttgct
     1741 ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa
     1801 aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac
     1861 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     1921 ggggtgttta ctagaggttg atcgggcacg taagaggttc caactttcac cataatgaaa
     1981 taagatcact accgggcgta ttttttgagt tatcgagatt ttcaggagct aaggaagcta
     2041 aaatgaggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     2101 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     2161 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
     2221 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     2281 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     2341 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     2401 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     2461 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     2521 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     2581 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     2641 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     2701 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     2761 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
     2821 tagtcggcaa ataatactag ctccggcaaa aaaacgggca aggtgtcacc accctgccct
     2881 ttttctttaa aaccgaaaag attacttcgc gtttgccacc tgacgtctaa gaaaaggaat
     2941 attcagcaat ttgcccgtgc cgaagaaagg cccacccgtg aaggtgagcc agtgagttga
     3001 ttgctacgta atgtcggcca attcgcgcta acttacatta attgcgttgc gctcactgcc
     3061 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3121 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagactggca
     3181 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     3241 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     3301 tatcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     3361 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     3421 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     3481 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     3541 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     3601 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcctcatgg gagaaaataa
     3661 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     3721 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     3781 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     3841 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     3901 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     3961 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     4021 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     4081 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcatattca
     4141 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4201 attcgatggc gcgccgcttc gtcaggccac atagctttct tgttctgatc ggaacgatcg
     4261 ttggctgtgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcgctcaca
     4321 attctatgga ctatgtttct gtcaccggat gtgctttccg gtctgatgag tccgtgagga
     4381 cgaaacagcc tctacaaata attttgttta atcacacagg aaagtactag
//
