LOCUS       sequence-95217-e        8438 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  beta core fragment expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  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 8438)
  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_42
FEATURES             Location/Qualifiers
     source          1..8438
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1817..1848
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             1934..1951
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2007..2024)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2177..2223
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      2279..2667
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(2717..3511)
                     /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     3438..3457
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      3917..4136
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             4227..4982
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    4985..5235
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             5561..6736
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             6736..7707
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    7780..8253
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     promoter        8377..8411
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
ORIGIN
        1 atgaataacg ctgccgccaa gcctctcatc actaccctac tccctaagat gattgcacgc
       61 atcaacgact ggtttgagga agtgaaagct aagcgcggca agcgcccgac agccttccag
      121 ttcctgcaag aaatcaagcc ggaagccgta gcgtacatca ccattaagac cactctggct
      181 tgcctaacca gtgctgacaa tacaaccgtt caggctgtag caagcgcaat cggtcgggcc
      241 attgaggacg aggctcgctt cggtcgtatc cgtgaccttg aagctaagca cttcaagaaa
      301 aacgttgagg aacaactcaa caagcgcgta gggcacgtct acaagaaagc atttatgcaa
      361 gttgtcgagg ctgacatgct ctctaagggt ctactcggtg gcgaggcgtg gtcttcgtgg
      421 cataaggaag actctattca tgtaggagta cgctgcatcg agatgctcat tgagtcaacc
      481 ggaatggtta gcttacaccg ccaaaatgct ggcgtagtag gtcaagactc tgagactatc
      541 gaactcgcac ctgaatacgc tgaggctatc gcaacccgtg caggtgcgct ggctggcatc
      601 tctccgatgt tccaaccttg cgtagttcct cctaagccgt ggactggcat tactggtggt
      661 ggctattggg ctaacggtcg tcgtcctctg gcgctggtgc gtactcacag taagaaagca
      721 ctgatgcgct acgaagacgt ttacatgcct gaggtgtaca aagcgattaa cattgcgcaa
      781 aacaccgcat ggaaaatcaa caagaaagtc ctagcggtcg ccaacgtaat caccaagtgg
      841 aagcattgtc cggtcgagga catccctgcg attgagcgtg aagaactccc gatgaaaccg
      901 gaagacatcg acatgaatcc tgaggctctc accgcgtgga aacgtgctgc cgctgctgtg
      961 taccgcaagg acaaggctcg caagtctcgc cgtatcagcc ttgagttcat gcttgagcaa
     1021 gccaataagt ttgctaacca taaggccatc tggttccctt acaacatgga ctggcgcggt
     1081 cgtgtttacg ctgtgtcaat gttcaacccg caaggtaacg atatgaccaa aggactgctt
     1141 acgctggcga aaggtaaacc aatcggtaag gaaggttact actggctgaa aatccacggt
     1201 gcaaactgtg cgggtgtcga caaggttccg ttccctgagc gcatcaagtt cattgaggaa
     1261 aaccacgaga acatcatggc ttgcgctaag tctccactgg agaacacttg gtgggctgag
     1321 caagattctc cgttctgctt ccttgcgttc tgctttgagt acgctggggt acagcaccac
     1381 ggcctgagct ataactgctc ccttccgctg gcgtttgacg ggtcttgctc tggcatccag
     1441 cacttctccg cgatgctccg agatgaggta ggtggtcgcg cggttaactt gcttcctagt
     1501 gaaaccgttc aggacatcta cgggattgtt gctaagaaag tcaacgagat tctacaagca
     1561 gacgcaatca atgggaccga taacgaagta gttaccgtga ccgatgagaa cggaggttca
     1621 ggtggtggat ccaacgaaaa agaagaactg aaatccaaaa aagcggaact gcgcaaccgt
     1681 atcgaacagc tgaaacagaa acgtgaacaa ctgaagcaga aaatcgcgaa cctgcgtaaa
     1741 gaaatcgaag cttacaaata atactagtgc ggccgctgca ggagtcacta agggttagtt
     1801 agttagatta gcagaaagtc aaaagcctcc gaccggaggc ttttgactaa aacttccctt
     1861 ggggttatca ttggggctca ctcaaaggcg gtaataactc atctcagaag aggatctgaa
     1921 tagcgccgtc gaccatcatc atcatcatca ttgagtttaa acggccgcca aaacagccaa
     1981 gctggagaat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     2041 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     2101 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     2161 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     2221 tatctgttgt ttgtcggtga actaattctg attcgcacgg gcccatggct aattcccatg
     2281 tcagccgtta agtgttcctg tgtcactcaa aattgctttg agaggctcta agggcttctc
     2341 agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc
     2401 ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat
     2461 ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta
     2521 cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc
     2581 ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca
     2641 acaggttgaa ctgctgatct tcagatcctc tacgccggac gcatcgtggc cggatcttgc
     2701 gcaagcggtc tccaattcag aagaactcgt caagaaggcg atagaaggcg atgcgctgcg
     2761 aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg ccgccaagct
     2821 cttcagcaat atcacgggta tccaacgcta tgtcctgata gcggtccgcc acacccagcc
     2881 ggccacagtc gttgaatcca gaaaagcggc cattttccac catgatattc ggcaagcagg
     2941 catcgccatg gatcacgacg agatcctcgc cgtcgggcat gcgcgccttg agcctggcga
     3001 acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga tcgacaagac
     3061 cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc
     3121 aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg gatactttct
     3181 cggcaggagc aaggcgagat gacaggagat cctgccccgg cacttcgccc aatagcagcc
     3241 agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg
     3301 ccagccacga tagccgcgct gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg
     3361 tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg gcatcagagc
     3421 agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa gcggccggag
     3481 aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct gtctcttgat
     3541 cagatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc cagtttactt
     3601 tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt tcgcttgctg
     3661 tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc
     3721 tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc atcaagctta
     3781 tgatgtggtc tgtcctttta cagccagtag tgctcgccgc agtcgagcga cagggcgaag
     3841 ccctcgagtg agcgaggaag caccagggaa cagcacttat atattctgct tacacacgat
     3901 gcctgaaaaa acttcccttg gggttatcca cttatccacg gggatatttt tataattatt
     3961 ttttttatag tttttagatc ttctttttta gagcgccttg taggccttta tccatgctgg
     4021 ttctagagaa ggtgttgtga caaattgccc tttcagtgtg acaaatcacc ctcaaatgac
     4081 agtcctgtct gtgacaaatt gcccttaacc ctgtgacaaa ttgccctcag aagaagctgt
     4141 tttttcacaa agttatccct gcttattgac tcttttttat ttagtgtgac aatctaaaaa
     4201 cttgtcacac ttcacatgga tctgtcatgg cggaaacagc ggttatcaat cacaagaaac
     4261 gtaaaaatag cccgcgaatc gtccagtcaa acgacctcac tgaggcggca tatagtctct
     4321 cccgggatca aaaacgtatg ctgtatctgt tcgttgacca gatcagaaaa tctgatggca
     4381 ccctacagga acatgacggt atctgcgaga tccatgttgc taaatatgct gaaatattcg
     4441 gattgacctc tgcggaagcc agtaaggata tacggcaggc attgaagagt ttcgcgggga
     4501 aggaagtggt tttttatcgc cctgaagagg atgccggcga tgaaaaaggc tatgaatctt
     4561 ttccttggtt tatcaaacgt gcgcacagtc catccagagg gctttacagt gtacatatca
     4621 acccatatct cattcccttc tttatcgggt tacagaaccg gtttacgcag tttcggctta
     4681 gtgaaacaaa agaaatcacc aatccgtatg ccatgcgttt atacgaatcc ctgtgtcagt
     4741 atcgtaagcc ggatggctca ggcatcgtct ctctgaaaat cgactggatc atagagcgtt
     4801 accagctgcc tcaaagttac cagcgtatgc ctgacttccg ccgccgcttc ctgcaggtct
     4861 gtgttaatga gatcaacagc agaactccaa tgcgcctctc atacattgag aaaaagaaag
     4921 gccgccagac gactcatatc gtattttcct tccgcgatat cacttccatg acgacaggat
     4981 agtctgaggg ttatctgtca cagatttgag ggtggttcgt cacatttgtt ctgacctact
     5041 gagggtaatt tgtcacagtt ttgctgtttc cttcagcctg catggatttt ctcatacttt
     5101 ttgaactgta atttttaagg aagccaaatt tgagggcagt ttgtcacagt tgatttcctt
     5161 ctctttccct tcgtcatgtg acctgatatc gggggttagt tcgtcatcat tgatgagggt
     5221 tgattatcac agtttattac tctgaattgg ctatccgcgt gtgtacctct acctggagtt
     5281 tttcccacgg tggatatttc ttcttgcgct gagcgtaaga gctatctgac agaacagttc
     5341 ttctttgctt cctcgccagt tcgctcgcta tgctcggtta cacggctgcg gcgagcgcta
     5401 gtgataataa gtgactgagg tatgtgctct tcttatctcc ttttgtagtg ttgctcttat
     5461 tttaaacaac tttgcggttt tttgatgact ttgcgatttt gttgttgctt tgcagtaaat
     5521 tgcaagattt aataaaaaaa cgcaaagcaa tgattaaagg atgttcagaa tgaaactcat
     5581 ggaaacactt aaccagtgca taaacgctgg tcatgaaatg acgaaggcta tcgccattgc
     5641 acagtttaat gatgacagcc cggaagcgag gaaaataacc cggcgctgga gaataggtga
     5701 agcagcggat ttagttgggg tttcttctca ggctatcaga gatgccgaga aagcagggcg
     5761 actaccgcac ccggatatgg aaattcgagg acgggttgag caacgtgttg gttatacaat
     5821 tgaacaaatt aatcatatgc gtgatgtgtt tggtacgcga ttgcgacgtg ctgaagacgt
     5881 atttccaccg gtgatcgggg ttgctgccca taaaggtggc gtttacaaaa cctcagtttc
     5941 tgttcatctt gctcaggatc tggctctgaa ggggctacgt gttttgctcg tggaaggtaa
     6001 cgacccccag ggaacagcct caatgtatca cggatgggta ccagatcttc atattcatgc
     6061 agaagacact ctcctgcctt tctatcttgg ggaaaaggac gatgtcactt atgcaataaa
     6121 gcccacttgc tggccggggc ttgacattat tccttcctgt ctggctctgc accgtattga
     6181 aactgagtta atgggcaaat ttgatgaagg taaactgccc accgatccac acctgatgct
     6241 ccgactggcc attgaaactg ttgctcatga ctatgatgtc atagttattg acagcgcgcc
     6301 taacctgggt atcggcacga ttaatgtcgt atgtgctgct gatgtgctga ttgttcccac
     6361 gcctgctgag ttgtttgact acacctccgc actgcagttt ttcgatatgc ttcgtgatct
     6421 gctcaagaac gttgatctta aagggttcga gcctgatgta cgtattttgc ttaccaaata
     6481 cagcaatagt aatggctctc agtccccgtg gatggaggag caaattcggg atgcctgggg
     6541 aagcatggtt ctaaaaaatg ttgtacgtga aacggatgaa gttggtaaag gtcagatccg
     6601 gatgagaact gtttttgaac aggccattga tcaacgctct tcaactggtg cctggagaaa
     6661 tgctctttct atttgggaac ctgtctgcaa tgaaattttc gatcgtctga ttaaaccacg
     6721 ctgggagatt agataatgaa gcgtgcgcct gttattccaa aacatacgct caatactcaa
     6781 ccggttgaag atacttcgtt atcgacacca gctgccccga tggtggattc gttaattgcg
     6841 cgcgtaggag taatggctcg cggtaatgcc attactttgc ctgtatgtgg tcgggatgtg
     6901 aagtttactc ttgaagtgct ccggggtgat agtgttgaga agacctctcg ggtatggtca
     6961 ggtaatgaac gtgaccagga gctgcttact gaggacgcac tggatgatct catcccttct
     7021 tttctactga ctggtcaaca gacaccggcg ttcggtcgaa gagtatctgg tgtcatagaa
     7081 attgccgatg ggagtcgccg tcgtaaagct gctgcactta ccgaaagtga ttatcgtgtt
     7141 ctggttggcg agctggatga tgagcagatg gctgcattat ccagattggg taacgattat
     7201 cgcccaacaa gtgcttatga acgtggtcag cgttatgcaa gccgattgca gaatgaattt
     7261 gctggaaata tttctgcgct ggctgatgcg gaaaatattt cacgtaagat tattacccgc
     7321 tgtatcaaca ccgccaaatt gcctaaatca gttgttgctc ttttttctca ccccggtgaa
     7381 ctatctgccc ggtcaggtga tgcacttcaa aaagccttta cagataaaga ggaattactt
     7441 aagcagcagg catctaacct tcatgagcag aaaaaagctg gggtgatatt tgaagctgaa
     7501 gaagttatca ctcttttaac ttctgtgctt aaaacgtcat ctgcatcaag aactagctta
     7561 agctcacgac atcagtttgc tcctggagcg acagtattgt ataagggcga taaaatggtg
     7621 cttaacctgg acaggtctcg tgttccaact gagtgtatag agaaaattga ggccattctt
     7681 aaggaacttg aaaagccagc accctgatgc gaccacgttt tagtctacgt ttatctgtct
     7741 ttacttaatg tcctttgtta caggccagaa agcataactg gcctgaatat tctctctggg
     7801 cccactgttc cacttgtatc gtcggtctga taatcagact gggaccacgg tcccactcgt
     7861 atcgtcggtc tgattattag tctgggacca cggtcccact cgtatcgtcg gtctgattat
     7921 tagtctggga ccacggtccc actcgtatcg tcggtctgat aatcagactg ggaccacggt
     7981 cccactcgta tcgtcggtct gattattagt ctgggaccat ggtcccactc gtatcgtcgg
     8041 tctgattatt agtctgggac cacggtccca ctcgtatcgt cggtctgatt attagtctgg
     8101 aaccacggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg
     8161 tatcgtcggt ctgattatta gtctgggacc acgatcccac tcgtgttgtc ggtctgatta
     8221 tcggtctggg accacggtcc cacttgtatt gtcgatcaga ctatcagcgt gagactacga
     8281 ttccatcaat gcctgtcaag ggcaagtatt gacatgtcgt cgtaacctgt agaacggagt
     8341 aacctcggtg tgcggttgta tgcctgctgt ggattgttta cggctagctc agtcctaggt
     8401 actatgctag ctactagagt cacacaggaa agtactag
//
