LOCUS       sequence-413708-        8810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7RNAP protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8810)
  TITLE     Patel lab plasmids
REFERENCE   2  (bases 1 to 8810)
  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"
COMMENT     Sequence Label: T7RNAP Cyslight A188C
FEATURES             Location/Qualifiers
     source          1..8810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(21..26,27..43,44..49)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(3094..3543)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGI"
     primer_bind     3492..3514
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3506..3523
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3507..3523
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3539..3555)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3539..3555)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3552..3574)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3563..3579
                     /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)."
     promoter        complement(join(3587..3593,3594..3611,3612..3617))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             complement(3666..4748)
                     /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     4710..4729
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4749..4826)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(4898..4917)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             6375..6566
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6560..6582)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6668..6808
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6723..6742
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6823..6840)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6994..7582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7074..7093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7753..8544,8545..8613))
                     /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     8376..8395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8614..8718)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8786..8804
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gtcttcaaga attctcatgt ttgacagctt atcatcgata agctttaatg cggtagttta
       61 tcacagttaa attgctaacg cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc
      121 gtcatcctcg gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg
      181 ccgggcctct tgcgggatat cgtccattcc gacagcatcg ccagtcacta tggcgtgctg
      241 ctagcgctat atgcgttgat gcaatttcta tgcgcacccg ttctcggagc actgtccgac
      301 cgctttggcc gccgcccagt cctgctcgct tcgctacttg gagccactat cgactacgcg
      361 atcatggcga ccacacccgt cctgtggatc cggagtcgta ttgatttggc gttacgcgaa
      421 cgcgaagtcc gactctaaga tgtcacggag gttcaagtta cctttagccg gaagtgctgg
      481 cattttgtcc aattgagact cgtgcaactg gtcagcgaac tggtcgtaga aatcagccag
      541 tacatcgcta gactcatatg tgtcaaccat agtttcgcgc actgctttga acaggttcgc
      601 agcgtcagcc ggaatggtac cgaaggagtc gtgaatcagt gcaaaagatt cgattccgta
      661 cttctcgtgt gcccacacta cagtcttacg aaggtggcta ccgtcttggc tgtgtacaaa
      721 gttaggagcg ataccagact cctgtttgtg tgcatcaatc tcgctatctt tgttggtgtt
      781 aatggtaggc tgtaagcgga actgaccgag gaacatcagg ttcaagcgcg tctgaatagg
      841 cttcttgtat tcctgccaca cagggaaacc atcaggagtt acccaatgca cagcgctacg
      901 cttgcgaaga atctctccag tcttcttatc tttgacctca gcagccagca gcttagcagc
      961 agacttaagc cagttcattg cttcaaccgc agctaccacc gtcacgctca cagattccca
     1021 aatcagctta gccatgtatc cagcagcctg attcggctga gtgaacatca gacccttgcc
     1081 ggaatcaata gctggctgaa tggtatcttc cagcacttgt tgacggaagc cgaactcttt
     1141 ggacccgtaa gccagcgtca tgactgaacg cttagtcaca ctgcgagtaa caccgtaagc
     1201 cagccattga ccagccagtg ccttagtgcc cagcttgact ttctcagaga tttcaccagt
     1261 gttctcatcg gtcacggtaa ctacttcgtt atcggtccca ttgattgcgt ctgcttgtag
     1321 aatctcgttg actttcttag caacaatccc gtagatgtcc tgaacggttt cactaggaag
     1381 caagttaacc gcgcgaccac ctacctcatc tcggagcatc gcggagaagt gctggatgcc
     1441 agagcaagac ccgtcaaacg ccagcggaag ggagctgtta tagctcaggc cgtggtgctg
     1501 taccccagcg tactcaaagc tgaacgcaag gaagctgaac ggagaatctt gctcagccca
     1561 ccaagtgttc tccagtggag acttagcgca agccatgatg ttctcgtggt tttcctcaat
     1621 gaacttgatg cgctcaggga acggaacctt atcgacaccc gcacagtttg caccgtggat
     1681 tttcagccag tagtaacctt ccttaccgat tggtttacct ttcgccagcg taagcagtcc
     1741 tttggtcata tcgttacctt gcgggttgaa cattgacaca gcgtaaacac gaccgcgcca
     1801 gtccatgttg taagggaacc agatggcctt atggttagca aacttattgg cttgctcaag
     1861 catgaactca aggctgatac ggcgagactt gcgagccttg tccttgcggt acacagcagc
     1921 ggcagcacgt ttccacgcgg tgagagcctc aggattcatg tcgatgtctt ccggtttcat
     1981 cgggagttct tcacgctcaa tcgcagggat gtcctcgacc gggctatgct tccacttggt
     2041 gattacgttg gcgaccgcta ggactttctt gttgattttc catgcggtgt tttgcgcaat
     2101 gttaatcgct ttgtacacct caggcatgta aacgtcttcg tagcgcatca gtgctttctt
     2161 actgtgagta cgcaccagcg ccagaggacg acgaccgtta gcccaatagc caccaccagt
     2221 aatgccagtc cacggcttag gaggaactac gcaaggttgg aacatcggag agatgccagc
     2281 cagcgcacct gcacgggttg cgatagcctc agcgtattca ggtgcgagtt cgatagtctc
     2341 agagtcttga cctactacgc cagcattttg gcggtgtaag ctaaccattc cggttgactc
     2401 aatgagcatc tcgatgcagc gtactcctac atgaatagag tcttccttat gccacgaaga
     2461 ccacgcctcg ccaccgagta gacccttaga gagcatgtcg cactcgacaa cttgcataaa
     2521 tgctttcttg tagacgtgcc ctacgcgctt gttgagttgt tcctcaacgt ttttcttgaa
     2581 gtgcttagct tcaaggtcac ggatacgacc gaagcgagcc tcgtcctcaa tggcccgacc
     2641 gattgcgctt gctacagcct gaacggttgt attgtcagca ctggttaggc tagccagagt
     2701 ggtcttaatg gtgatgtacg ctacggcttc cggcttgatt tcttgcagga actggaaggc
     2761 tgtcgggcgc ttgccgcgct tagctttcac ttcctcaaac cagtcgttga tgcgtgcaat
     2821 catcttaggg agtagggtag tgatgagagg cttggcggca gcgttatccg caacctcacc
     2881 agctttaagt tgacgctcaa acatcttgcg gaagcgtgct tcacccatct cgtaagactc
     2941 atgctcaagg gccaactgtt cgcgagctaa acgctcaccg taatggtcag ccagagtgtt
     3001 gaacgggata gcagccagtt cgatgtcaga gaagtcgttc ttagcgatgt taatcgtgtt
     3061 catttagtgc ctcttccagt tagtaaatcc ggatcagatc ccgacgccat caaaaataat
     3121 tcgcgtctgg ccttcctgta gccagctttc atcaacatta aatgtgagcg agtaacaacc
     3181 cgtcggattc tccgtgggaa caaacggcgg attgaccgta atgggatagg tcacgttggt
     3241 gtagatgggc gcatcgtaac cgtgcatctg ccagtttgag gggacgacga cagtatcggc
     3301 ctcaggaaga tcgcactcca gccagctttc cggcaccgct tctggtgccg gaaaccaggc
     3361 aaagcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     3421 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     3481 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaatccg taatcatggt
     3541 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacatt atacgagccg
     3601 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaacttaca ttaattgcgt
     3661 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     3721 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc
     3781 agtgagacgg gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag
     3841 cggtccacgc tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg
     3901 atataacatg agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg
     3961 cgcagcccgg actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc
     4021 agcatcgcag tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac
     4081 atggcactcc agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat
     4141 ttatgccagc cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc
     4201 gcgatttgct ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca
     4261 tgggagaaaa taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga
     4321 acattagtgc aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg
     4381 atcagcccac tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg
     4441 ccgcttcgtt ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta
     4501 atcgccgcga caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc
     4561 agcaacgact gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc
     4621 gccatcgccg cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc
     4681 acgcgggaaa cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact
     4741 ggtttcacat tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga
     4801 aaggttttgc gccattcgat ggtgtccggg atctggatcc tctacgccgg acgcatcgtg
     4861 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     4921 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     4981 gcaggccccg tggccggggg actgttgggc gccatctcct tgcatgcacc attccttgcg
     5041 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     5101 aagggagagc gtcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     5161 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     5221 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     5281 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     5341 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     5401 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc
     5461 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc
     5521 aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc
     5581 ctaacttcga tcattggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca
     5641 tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg
     5701 cgtcgcggtg catggagccg ggccacctcg acctgaatgg aagccggcgg cacctcgcta
     5761 acggattcac cactccaaga attggagcca atcaattctt gcggagaact gtgaatgcgc
     5821 aaaccaaccc ttggcagaac atatccatcg cgtccgccat ctccagcagc cgcacgcggc
     5881 gcatctcggg cagcgttggg tcctggccac gggtgcgcat gatcgtgctc ctgtcgttga
     5941 ggacccggct aggctggcgg ggttgcctta ctggttagca gaatgaatca ccgatacgcg
     6001 agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg
     6061 tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta
     6121 tgttccggat ctgcatcgca ggatgctgct ggctaccctg tggaacacct acatctgtat
     6181 taacgaagcg ctggcattga ccctgagtga tttttctctg gtcccgccgc atccataccg
     6241 ccagttgttt accctcacaa cgttccagta accgggcatg ttcatcatca gtaacccgta
     6301 tcgtgagcat cctctctcgt ttcatcggta tcattacccc catgaacaga aatccccctt
     6361 acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc cgctttatca
     6421 gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat gaacaggcag
     6481 acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt
     6541 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     6601 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     6661 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
     6721 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     6781 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
     6841 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     6901 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     6961 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7021 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7081 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7141 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7201 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7261 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7321 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7381 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7441 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7501 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7561 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7621 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7681 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7741 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7801 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7861 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7921 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7981 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8041 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     8101 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8161 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8221 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8281 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8341 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     8401 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8461 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8521 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8581 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8641 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8701 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     8761 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
//
