LOCUS       sequence-95215-e        9330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  High full length T7 RNAP* expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9330)
  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 9330)
  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_41
FEATURES             Location/Qualifiers
     source          1..9330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    2653..2673
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      2709..2740
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             2826..2843
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2899..2916)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3069..3115
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      3171..3559
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(3609..4403)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     4330..4349
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      4809..5028
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             5119..5874
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    5877..6127
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             6453..7628
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             7628..8599
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    8672..9145
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     promoter        9269..9303
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
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 cgctgccgcc aagcctctca tcactaccct actccctaag
      241 atgattgcac gcatcaacga ctggtttgag gaagtgaaag ctaagcgcgg caagcgcccg
      301 acagccttcc agttcctgca agaaatcaag ccggaagccg tagcgtacat caccattaag
      361 accactctgg cttgcctaac cagtgctgac aatacaaccg ttcaggctgt agcaagcgca
      421 atcggtcggg ccattgagga cgaggctcgc ttcggtcgta tccgtgacct tgaagctaag
      481 cacttcaaga aaaacgttga ggaacaactc aacaagcgcg tagggcacgt ctacaagaaa
      541 gcatttatgc aagttgtcga ggctgacatg ctctctaagg gtctactcgg tggcgaggcg
      601 tggtcttcgt ggcataagga agactctatt catgtaggag tacgctgcat cgagatgctc
      661 attgagtcaa ccggaatggt tagcttacac cgccaaaatg ctggcgtagt aggtcaagac
      721 tctgagacta tcgaactcgc acctgaatac gctgaggcta tcgcaacccg tgcaggtgcg
      781 ctggctggca tctctccgat gttccaacct tgcgtagttc ctcctaagcc gtggactggc
      841 attactggtg gtggctattg ggctaacggt cgtcgtcctc tggcgctggt gcgtactcac
      901 agtaagaaag cactgatgcg ctacgaagac gtttacatgc ctgaggtgta caaagcgatt
      961 aacattgcgc aaaacaccgc atggaaaatc aacaagaaag tcctagcggt cgccaacgta
     1021 atcaccaagt ggaagcattg tccggtcgag gacatccctg cgattgagcg tgaagaactc
     1081 ccgatgaaac cggaagacat cgacatgaat cctgaggctc tcaccgcgtg gaaacgtgct
     1141 gccgctgctg tgtaccgcaa ggacaaggct cgcaagtctc gccgtatcag ccttgagttc
     1201 atgcttgagc aagccaataa gtttgctaac cataaggcca tctggttccc ttacaacatg
     1261 gactggcgcg gtcgtgttta cgctgtgtca atgttcaacc cgcaaggtaa cgatatgacc
     1321 aaaggactgc ttacgctggc gaaaggtaaa ccaatcggta aggaaggtta ctactggctg
     1381 aaaatccacg gtgcaaactg tgcgggtgtc gacaaggttc cgttccctga gcgcatcaag
     1441 ttcattgagg aaaaccacga gaacatcatg gcttgcgcta agtctccact ggagaacact
     1501 tggtgggctg agcaagattc tccgttctgc ttccttgcgt tctgctttga gtacgctggg
     1561 gtacagcacc acggcctgag ctataactgc tcccttccgc tggcgtttga cgggtcttgc
     1621 tctggcatcc agcacttctc cgcgatgctc cgagatgagg taggtggtcg cgcggttaac
     1681 ttgcttccta gtgaaaccgt tcaggacatc tacgggattg ttgctaagaa agtcaacgag
     1741 attctacaag cagacgcaat caatgggacc gataacgaag tagttaccgt gaccgatgag
     1801 aacactggtg aaatctctga gaaagtcaag ctgggcacta aggcactggc tggtcaatgg
     1861 ctggcttacg gtgttactcg cagtgtgact aagagttcag tcatgacgct ggcttacggg
     1921 tccaaagagt tcggcttccg tcaacaagtg ctggaagata ccattcagcc agctattgat
     1981 tccggcaagg gtctgatgtt cactcagccg aatcaggctg ctggatacat ggctaagctg
     2041 atttgggaat ctgtgagcgt gacggtggta gctgcggttg aagcaatgaa ctggcttaag
     2101 tctgctgcta agctgctggc tgctgaggtc aaagataaga agactggaga gattcttcgc
     2161 aagcgttgcg ctgtgcattg ggtaactcct gatggtttcc ctgtgtggca ggaatacaag
     2221 aagcctattc agacgcgctt gaacctgatg ttcctcggtc agttccgctt acagcctacc
     2281 attaacacca acaaagatag cgagattgat gcacacaaac aggagtctgg tatcgctcct
     2341 aactttgtac acagccaaga cggtagccac cttcgtaaga ctgtagtgtg ggcacacgag
     2401 aagtacggaa tcgaatcttt tgcactgatt cacgactcct tcggtacgat tccggctgac
     2461 gctgcgaacc tgttcaaagc agtgcgcgaa actatggttg acacatatga gtcttgtgat
     2521 gtactggctg atttctacga ccagttcgct gaccagttgc acgagtctca attggacaaa
     2581 atgccagcac ttccggctaa aggtaacttg aacctccgtg acatcttaga gtcggacttc
     2641 gcgttcgcgt aatactagta gcggccgctg caggagtcac taagggttag ttagttagat
     2701 tagcagaaag tcaaaagcct ccgaccggag gcttttgact aaaacttccc ttggggttat
     2761 cattggggct cactcaaagg cggtaataac tcatctcaga agaggatctg aatagcgccg
     2821 tcgaccatca tcatcatcat cattgagttt aaacggccgc caaaacagcc aagctggaga
     2881 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     2941 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     3001 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     3061 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     3121 gtttgtcggt gaactaattc tgattcgcac gggcccatgg ctaattccca tgtcagccgt
     3181 taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc tcagtgcgtt
     3241 acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag ccttatatat
     3301 tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg atttatatta
     3361 attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag tacgttagcc
     3421 atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga gcttagtacg
     3481 ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat caacaggttg
     3541 aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatctt gcgcaagcgg
     3601 tctccaattc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     3661 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     3721 atatcacggg tatccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     3781 tcgttgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     3841 tggatcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg
     3901 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
     3961 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
     4021 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
     4081 gcaaggcgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
     4141 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
     4201 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca
     4261 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
     4321 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
     4381 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatctt
     4441 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
     4501 ttcccaacct taccagaggg cgccccagct ggcaattccg gttcgcttgc tgtccataaa
     4561 accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg
     4621 cttgcgtttt cccttgtcca gatagcccag tagctgacat tcatcaagct tatgatgtgg
     4681 tctgtccttt tacagccagt agtgctcgcc gcagtcgagc gacagggcga agccctcgag
     4741 tgagcgagga agcaccaggg aacagcactt atatattctg cttacacacg atgcctgaaa
     4801 aaacttccct tggggttatc cacttatcca cggggatatt tttataatta ttttttttat
     4861 agtttttaga tcttcttttt tagagcgcct tgtaggcctt tatccatgct ggttctagag
     4921 aaggtgttgt gacaaattgc cctttcagtg tgacaaatca ccctcaaatg acagtcctgt
     4981 ctgtgacaaa ttgcccttaa ccctgtgaca aattgccctc agaagaagct gttttttcac
     5041 aaagttatcc ctgcttattg actctttttt atttagtgtg acaatctaaa aacttgtcac
     5101 acttcacatg gatctgtcat ggcggaaaca gcggttatca atcacaagaa acgtaaaaat
     5161 agcccgcgaa tcgtccagtc aaacgacctc actgaggcgg catatagtct ctcccgggat
     5221 caaaaacgta tgctgtatct gttcgttgac cagatcagaa aatctgatgg caccctacag
     5281 gaacatgacg gtatctgcga gatccatgtt gctaaatatg ctgaaatatt cggattgacc
     5341 tctgcggaag ccagtaagga tatacggcag gcattgaaga gtttcgcggg gaaggaagtg
     5401 gttttttatc gccctgaaga ggatgccggc gatgaaaaag gctatgaatc ttttccttgg
     5461 tttatcaaac gtgcgcacag tccatccaga gggctttaca gtgtacatat caacccatat
     5521 ctcattccct tctttatcgg gttacagaac cggtttacgc agtttcggct tagtgaaaca
     5581 aaagaaatca ccaatccgta tgccatgcgt ttatacgaat ccctgtgtca gtatcgtaag
     5641 ccggatggct caggcatcgt ctctctgaaa atcgactgga tcatagagcg ttaccagctg
     5701 cctcaaagtt accagcgtat gcctgacttc cgccgccgct tcctgcaggt ctgtgttaat
     5761 gagatcaaca gcagaactcc aatgcgcctc tcatacattg agaaaaagaa aggccgccag
     5821 acgactcata tcgtattttc cttccgcgat atcacttcca tgacgacagg atagtctgag
     5881 ggttatctgt cacagatttg agggtggttc gtcacatttg ttctgaccta ctgagggtaa
     5941 tttgtcacag ttttgctgtt tccttcagcc tgcatggatt ttctcatact ttttgaactg
     6001 taatttttaa ggaagccaaa tttgagggca gtttgtcaca gttgatttcc ttctctttcc
     6061 cttcgtcatg tgacctgata tcgggggtta gttcgtcatc attgatgagg gttgattatc
     6121 acagtttatt actctgaatt ggctatccgc gtgtgtacct ctacctggag tttttcccac
     6181 ggtggatatt tcttcttgcg ctgagcgtaa gagctatctg acagaacagt tcttctttgc
     6241 ttcctcgcca gttcgctcgc tatgctcggt tacacggctg cggcgagcgc tagtgataat
     6301 aagtgactga ggtatgtgct cttcttatct ccttttgtag tgttgctctt attttaaaca
     6361 actttgcggt tttttgatga ctttgcgatt ttgttgttgc tttgcagtaa attgcaagat
     6421 ttaataaaaa aacgcaaagc aatgattaaa ggatgttcag aatgaaactc atggaaacac
     6481 ttaaccagtg cataaacgct ggtcatgaaa tgacgaaggc tatcgccatt gcacagttta
     6541 atgatgacag cccggaagcg aggaaaataa cccggcgctg gagaataggt gaagcagcgg
     6601 atttagttgg ggtttcttct caggctatca gagatgccga gaaagcaggg cgactaccgc
     6661 acccggatat ggaaattcga ggacgggttg agcaacgtgt tggttataca attgaacaaa
     6721 ttaatcatat gcgtgatgtg tttggtacgc gattgcgacg tgctgaagac gtatttccac
     6781 cggtgatcgg ggttgctgcc cataaaggtg gcgtttacaa aacctcagtt tctgttcatc
     6841 ttgctcagga tctggctctg aaggggctac gtgttttgct cgtggaaggt aacgaccccc
     6901 agggaacagc ctcaatgtat cacggatggg taccagatct tcatattcat gcagaagaca
     6961 ctctcctgcc tttctatctt ggggaaaagg acgatgtcac ttatgcaata aagcccactt
     7021 gctggccggg gcttgacatt attccttcct gtctggctct gcaccgtatt gaaactgagt
     7081 taatgggcaa atttgatgaa ggtaaactgc ccaccgatcc acacctgatg ctccgactgg
     7141 ccattgaaac tgttgctcat gactatgatg tcatagttat tgacagcgcg cctaacctgg
     7201 gtatcggcac gattaatgtc gtatgtgctg ctgatgtgct gattgttccc acgcctgctg
     7261 agttgtttga ctacacctcc gcactgcagt ttttcgatat gcttcgtgat ctgctcaaga
     7321 acgttgatct taaagggttc gagcctgatg tacgtatttt gcttaccaaa tacagcaata
     7381 gtaatggctc tcagtccccg tggatggagg agcaaattcg ggatgcctgg ggaagcatgg
     7441 ttctaaaaaa tgttgtacgt gaaacggatg aagttggtaa aggtcagatc cggatgagaa
     7501 ctgtttttga acaggccatt gatcaacgct cttcaactgg tgcctggaga aatgctcttt
     7561 ctatttggga acctgtctgc aatgaaattt tcgatcgtct gattaaacca cgctgggaga
     7621 ttagataatg aagcgtgcgc ctgttattcc aaaacatacg ctcaatactc aaccggttga
     7681 agatacttcg ttatcgacac cagctgcccc gatggtggat tcgttaattg cgcgcgtagg
     7741 agtaatggct cgcggtaatg ccattacttt gcctgtatgt ggtcgggatg tgaagtttac
     7801 tcttgaagtg ctccggggtg atagtgttga gaagacctct cgggtatggt caggtaatga
     7861 acgtgaccag gagctgctta ctgaggacgc actggatgat ctcatccctt cttttctact
     7921 gactggtcaa cagacaccgg cgttcggtcg aagagtatct ggtgtcatag aaattgccga
     7981 tgggagtcgc cgtcgtaaag ctgctgcact taccgaaagt gattatcgtg ttctggttgg
     8041 cgagctggat gatgagcaga tggctgcatt atccagattg ggtaacgatt atcgcccaac
     8101 aagtgcttat gaacgtggtc agcgttatgc aagccgattg cagaatgaat ttgctggaaa
     8161 tatttctgcg ctggctgatg cggaaaatat ttcacgtaag attattaccc gctgtatcaa
     8221 caccgccaaa ttgcctaaat cagttgttgc tcttttttct caccccggtg aactatctgc
     8281 ccggtcaggt gatgcacttc aaaaagcctt tacagataaa gaggaattac ttaagcagca
     8341 ggcatctaac cttcatgagc agaaaaaagc tggggtgata tttgaagctg aagaagttat
     8401 cactctttta acttctgtgc ttaaaacgtc atctgcatca agaactagct taagctcacg
     8461 acatcagttt gctcctggag cgacagtatt gtataagggc gataaaatgg tgcttaacct
     8521 ggacaggtct cgtgttccaa ctgagtgtat agagaaaatt gaggccattc ttaaggaact
     8581 tgaaaagcca gcaccctgat gcgaccacgt tttagtctac gtttatctgt ctttacttaa
     8641 tgtcctttgt tacaggccag aaagcataac tggcctgaat attctctctg ggcccactgt
     8701 tccacttgta tcgtcggtct gataatcaga ctgggaccac ggtcccactc gtatcgtcgg
     8761 tctgattatt agtctgggac cacggtccca ctcgtatcgt cggtctgatt attagtctgg
     8821 gaccacggtc ccactcgtat cgtcggtctg ataatcagac tgggaccacg gtcccactcg
     8881 tatcgtcggt ctgattatta gtctgggacc atggtcccac tcgtatcgtc ggtctgatta
     8941 ttagtctggg accacggtcc cactcgtatc gtcggtctga ttattagtct ggaaccacgg
     9001 tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg
     9061 gtctgattat tagtctggga ccacgatccc actcgtgttg tcggtctgat tatcggtctg
     9121 ggaccacggt cccacttgta ttgtcgatca gactatcagc gtgagactac gattccatca
     9181 atgcctgtca agggcaagta ttgacatgtc gtcgtaacct gtagaacgga gtaacctcgg
     9241 tgtgcggttg tatgcctgct gtggattgtt tacggctagc tcagtcctag gtactatgct
     9301 agctactaga gtcatttatg aaagtactag
//
