LOCUS       sequence-229806-        8621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 RNAP-driven expression of N-terminal His-tagged lacZ omega 
            subunit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8621)
  AUTHORS   Ma D, Shen L, Wu K, Diehnelt CW, Green AA
  TITLE     Low-cost detection of norovirus using paper-based cell-free systems 
            and synbody-based viral enrichment.
  JOURNAL   Synth Biol (Oxf). 2018;3(1):ysy018. doi: 10.1093/synbio/ysy018. Epub
            2018 Sep 19.
  PUBMED    30370338
REFERENCE   2  (bases 1 to 8621)
  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: "Synth Biol 
            (Oxf)."; date: "2018"; volume: "3(1):ysy018. doi"; pages: " 
            10.1093/synbio/ysy018. Epub 2018 Sep 19"
COMMENT     Sequence Label: pDM_T7_HisLacZomega
FEATURES             Location/Qualifiers
     source          1..8621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     terminator      3136..3183
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3358..3363,3364..3380,3381..3386))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3413..3431)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3499..3603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3604..3672,3673..4464)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3822..3841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4635..5223
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5124..5143
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5377..5394
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5409..5548
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5474..5493)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5634..5656
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5650..5841)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7153..8235)
                     /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     8197..8216
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8236..8313)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8519..8538
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg cgtaaaatga ccatgattac ggattcactg
      181 gccgtcgccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
      241 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
      301 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
      361 tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
      421 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
      481 cgaattattt ttgatggcgt taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc
      541 ggttacggcc aggacagtcg tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc
      601 ggagaaaacc gcctcgcggt gatggtgctg cgctggagtg acggcagtta tctggaagat
      661 caggatatgt ggcggatgag cggcattttc cgtgacgtct cgttgctgca taaaccgact
      721 acacaaatca gcgatttcca tgttgccact cgctttaatg atgatttcag ccgcgctgta
      781 ctggaggctg aagttcagat gtgcggcgag ttgcgtgact acctacgggt aacagtttct
      841 ttatggcagg gtgaaacgca ggtcgccagc ggcaccgcgc ctttcggcgg tgaaattatc
      901 gatgagcgtg gtggttatgc cgatcgcgtc acactacgtc tgaacgtcga aaacccgaaa
      961 ctgtggagcg ccgaaatccc gaatctctat cgtgcggtgg ttgaactgca caccgccgac
     1021 ggcacgctga ttgaagcaga agcctgcgat gtcggtttcc gcgaggtgcg gattgaaaat
     1081 ggtctgctgc tgctgaacgg caagccgttg ctgattcgag gcgttaaccg tcacgagcat
     1141 catcctctgc atggtcaggt catggatgag cagacgatgg tgcaggatat cctgctgatg
     1201 aagcagaaca actttaacgc cgtgcgctgt tcgcattatc cgaaccatcc gctgtggtac
     1261 acgctgtgcg accgctacgg cctgtatgtg gtggatgaag ccaatattga aacccacggc
     1321 atggtgccaa tgaatcgtct gaccgatgat ccgcgctggc taccggcgat gagcgaacgc
     1381 gtaacgcgaa tggtgcagcg cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg
     1441 aatgaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa atctgtcgat
     1501 ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac cgatattatt
     1561 tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc gaaatggtcc
     1621 atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgatcctttg cgaatacgcc
     1681 cacgcgatgg gtaacagtct tggcggtttc gctaaatact ggcaggcgtt tcgtcagtat
     1741 ccccgtttac agggcggctt cgtctgggac tgggtggatc agtcgctgat taaatatgat
     1801 gaaaacggca acccgtggtc ggcttacggc ggtgattttg gcgatacgcc gaacgatcgc
     1861 cagttctgta tgaacggtct ggtctttgcc gaccgcacgc cgcatccagc gctgacggaa
     1921 gcaaaacacc agcagcagtt tttccagttc cgtttatccg ggcaaaccat cgaagtgacc
     1981 agcgaatacc tgttccgtca tagcgataac gagctcctgc actggatggt ggcgctggat
     2041 ggtaagccgc tggcaagcgg tgaagtgcct ctggatgtcg ctccacaagg taaacagttg
     2101 attgaactgc ctgaactacc gcagccggag agcgccgggc aactctggct cacagtacgc
     2161 gtagtgcaac cgaacgcgac cgcatggtca gaagccgggc acatcagcgc ctggcagcag
     2221 tggcgtctgg cggaaaacct cagtgtgacg ctccccgccg cgtcccacgc catcccgcat
     2281 ctgaccacca gcgaaatgga tttttgcatc gagctgggta ataagcgttg gcaatttaac
     2341 cgccagtcag gctttctttc acagatgtgg attggcgata aaaaacaact gctgacgccg
     2401 ctgcgcgatc agttcacccg tgcaccgctg gataacgaca ttggcgtaag tgaagcgacc
     2461 cgcattgacc ctaacgcctg ggtcgaacgc tggaaggcgg cgggccatta ccaggccgaa
     2521 gcagcgttgt tgcagtgcac ggcagataca cttgctgatg cggtgctgat tacgaccgct
     2581 cacgcgtggc agcatcaggg gaaaacctta tttatcagcc ggaaaaccta ccggattgat
     2641 ggtagtggtc aaatggcgat taccgttgat gttgaagtgg cgagcgatac accgcatccg
     2701 gcgcggattg gcctgaactg ccagctggcg caggtagcag agcgggtaaa ctggctcgga
     2761 ttagggccgc aagaaaacta tcccgaccgc cttactgccg cctgttttga ccgctgggat
     2821 ctgccattgt cagacatgta taccccgtac gtcttcccga gcgaaaacgg tctgcgctgc
     2881 gggacgcgcg aattgaatta tggcccacac cagtggcgcg gcgacttcca gttcaacatc
     2941 agccgctaca gtcaacagca actgatggaa accagccatc gccatctgct gcacgcggaa
     3001 gaaggcacat ggctgaatat cgacggtttc catatgggga ttggtggcga cgactcctgg
     3061 agcccgtcag tatcggcgga attccagctg agcgccggtc gctaccatta ccagttggtc
     3121 tggtgtcaaa aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     3181 ttgctgaaag gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa
     3241 ccaagcctat gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt
     3301 tagatttcat acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt
     3361 aaagcttatc gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg
     3421 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     3481 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3541 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3601 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3661 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3721 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3781 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3841 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3901 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3961 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     4021 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4081 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4141 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4201 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4261 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4321 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4381 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4441 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     4501 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     4561 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     4621 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     4681 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4741 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     4801 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4861 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4921 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4981 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     5041 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     5101 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     5161 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     5221 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     5281 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     5341 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     5401 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     5461 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     5521 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     5581 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     5641 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     5701 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     5761 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     5821 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     5881 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     5941 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     6001 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     6061 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     6121 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     6181 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     6241 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     6301 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     6361 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     6421 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     6481 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     6541 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     6601 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     6661 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     6721 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     6781 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     6841 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     6901 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     6961 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     7021 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     7081 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     7141 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     7201 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     7261 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     7321 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     7381 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     7441 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     7501 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     7561 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     7621 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     7681 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     7741 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     7801 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     7861 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     7921 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     7981 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     8041 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     8101 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     8161 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     8221 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     8281 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     8341 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     8401 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     8461 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     8521 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     8581 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa t
//
