LOCUS       sequence-405325-        5659 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 5659)
  TITLE     Patel lab plasmids
REFERENCE   2  (bases 1 to 5659)
  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 V166C
FEATURES             Location/Qualifiers
     source          1..5659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             18..40
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(89..105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(89..105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(102..124)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    113..129
                     /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(137..143,144..161,162..167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    182..203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(288..310)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    396..536
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     451..470
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(551..568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(722..1310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(802..821)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1481..2272,2273..2341))
                     /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     2104..2123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2342..2446)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2514..2532
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(2559..2564,2565..2581,2582..2587)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(5614..5631)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(5641..5658)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ctgctgccca tggtatatct ccttcttaaa gttaaacaaa attatttcta gagtcgacct
       61 gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      121 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      181 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      241 acctgtcgtg ccagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
      301 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
      361 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
      421 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      481 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
      541 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
      601 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      661 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      721 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      781 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      841 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      901 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      961 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1021 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1081 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1141 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     1201 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     1261 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     1321 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     1381 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     1441 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     1501 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1561 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1621 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1681 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1741 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     1801 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1861 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1921 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1981 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2041 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     2101 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     2161 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     2221 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     2281 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     2341 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     2401 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     2461 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     2521 gtatcacgag gccctttcgt cttcaagaat tctcatgttt gacagcttat catcgataag
     2581 ctttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa
     2641 tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc
     2701 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc
     2761 agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt
     2821 ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga
     2881 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatccg gagtcgtatt
     2941 gatttggcgt tacgcgaacg cgaagtccga ctctaagatg tcacggaggt tcaagttacc
     3001 tttagccgga agtgctggca ttttgtccaa ttgagactcg tgcaactggt cagcgaactg
     3061 gtcgtagaaa tcagccagta catcgctaga ctcatatgtg tcaaccatag tttcgcgcac
     3121 tgctttgaac aggttcgcag cgtcagccgg aatggtaccg aaggagtcgt gaatcagtgc
     3181 aaaagattcg attccgtact tctcgtgtgc ccacactaca gtcttacgaa ggtggctacc
     3241 gtcttggctg tgtacaaagt taggagcgat accagactcc tgtttgtgtg catcaatctc
     3301 gctatctttg ttggtgttaa tggtaggctg taagcggaac tgaccgagga acatcaggtt
     3361 caagcgcgtc tgaataggct tcttgtattc ctgccacaca gggaaaccat caggagttac
     3421 ccaatgcaca gcgctacgct tgcgaagaat ctctccagtc ttcttatctt tgacctcagc
     3481 agccagcagc ttagcagcag acttaagcca gttcattgct tcaaccgcag ctaccaccgt
     3541 cacgctcaca gattcccaaa tcagcttagc catgtatcca gcagcctgat tcggctgagt
     3601 gaacatcaga cccttgccgg aatcaatagc tggctgaatg gtatcttcca gcacttgttg
     3661 acggaagccg aactctttgg acccgtaagc cagcgtcatg actgaacgct tagtcacact
     3721 gcgagtaaca ccgtaagcca gccattgacc agccagtgcc ttagtgccca gcttgacttt
     3781 ctcagagatt tcaccagtgt tctcatcggt cacggtaact acttcgttat cggtcccatt
     3841 gattgcgtct gcttgtagaa tctcgttgac tttcttagca acaatcccgt agatgtcctg
     3901 aacggtttca ctaggaagca agttaaccgc gcgaccacct acctcatctc ggagcatcgc
     3961 ggagaagtgc tggatgccag agcaagaccc gtcaaacgcc agcggaaggg agctgttata
     4021 gctcaggccg tggtgctgta ccccagcgta ctcaaagctg aacgcaagga agctgaacgg
     4081 agaatcttgc tcagcccacc aagtgttctc cagtggagac ttagcgcaag ccatgatgtt
     4141 ctcgtggttt tcctcaatga acttgatgcg ctcagggaac ggaaccttat cgacacccgc
     4201 acagtttgca ccgtggattt tcagccagta gtaaccttcc ttaccgattg gtttaccttt
     4261 cgccagcgta agcagtcctt tggtcatatc gttaccttgc gggttgaaca ttgacacagc
     4321 gtaaacacga ccgcgccagt ccatgttgta agggaaccag atggccttat ggttagcaaa
     4381 cttattggct tgctcaagca tgaactcaag gctgatacgg cgagacttgc gagccttgtc
     4441 cttgcggtac acagcagcgg cagcacgttt ccacgcggtg agagcctcag gattcatgtc
     4501 gatgtcttcc ggtttcatcg ggagttcttc acgctcaatc gcagggatgt cctcgaccgg
     4561 gctatgcttc cacttggtga ttacgttggc gaccgctagg actttcttgt tgattttcca
     4621 tgcggtgttt tgcgcaatgt taatcgcttt gtacacctca ggcatgtaaa cgtcttcgta
     4681 gcgcatcagt gctttcttac tgtgagtacg caccagcgcc agaggacgac gaccgttagc
     4741 ccaatagcca ccaccagtaa tgccagtcca cggcttagga ggaactacgc aaggttggaa
     4801 catcggagag atgccagcca gcgcacctgc acgggttgcg atagcctcag cgtattcagg
     4861 tgcgagttcg atagtctcag agtcttgacc tactacgcca gcattttggc ggtgtaagct
     4921 aaccattccg gttgactcaa tgagcatctc gatgcagcgt actcctacat gaatagagtc
     4981 ttccttatgc cacgaagacc acgcctcgcc accgagtaga cccttagaga gcatgtcagc
     5041 ctcgacaact tgcataaatg ctttcttgta gacgtgccct acgcgcttgt tgagttgttc
     5101 ctcaacgttt ttcttgaagt gcttagcttc aaggtcacgg atacgaccga agcgagcctc
     5161 gtcctcaatg gcccgaccga ttgcgcttgc tacagcctga acggttgtat tgtcagcact
     5221 ggttaggcta gccagagtgg tcttaatggt gatgtacgct acggcttccg gcttgatttc
     5281 ttgcaggaac tggaaggctg tcgggcgctt gccgcgctta gctttcactt cctcaaacca
     5341 gtcgttgatg cgtgcaatca tcttagggag tagggtagtg atgagaggct tggcggcagc
     5401 gttatccgca acctcaccag ctttaagttg acgctcaaac atcttgcgga agcgtgcttc
     5461 acccatctcg taagactcat gctcaagggc caactgttcg cgagctaaac gctcaccgta
     5521 atggtcagcc agagtgttga acgggatagc agccagttcg atgtcagaga agtcgttctt
     5581 agcgatgtta atcgtgttca tctcgagcat atggctgccg cgcggcacca ggccgctgct
     5641 gtgatgatga tgatgatgg
//
