LOCUS       sequence-296717-        6437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T7 promoter driven synthesis of T3 RNA polymerase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6437)
  AUTHORS   Niederholtmeyer H, Chaggan C, Devaraj NK
  TITLE     Communication and quorum sensing in non-living mimics of eukaryotic 
            cells.
  JOURNAL   Nat Commun. 2018 Nov 28;9(1):5027. doi: 10.1038/s41467-018-07473-7.
  PUBMED    30487584
REFERENCE   2  (bases 1 to 6437)
  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: "Nat 
            Commun."; date: "2018-11-28"; volume: "9(1):5027. doi"; pages: " 
            10.1038/s41467-018-07473-7"
COMMENT     Sequence Label: pSC101-pT7-T3RNAP
FEATURES             Location/Qualifiers
     source          1..6437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     113..132
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        113..131
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      2855..2902
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             3116..3910
                     /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
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3170..3189)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3780..3799
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      3941..4035
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4797..5747
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(6278..6305)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6321..6392)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccgagcgact ctcaaggtgc agacgtaaaa aaagcggcgt ggttagccgc ttttttaatt
       61 gccggaagtt cattaagctg ctgtcttcac ctcgaggatc ttaaggctag agtaatacga
      121 ctcactatag ggagatgtgg tctagacatt ccaggttaag aaggaggaaa aaaaatgaac
      181 atcatcgaaa acatcgaaaa gaatgacttc tcagaaatcg aactggctgc tatcccgttc
      241 aacacactgg ctgaccacta cggaagcgcc ttggctaaag agcagttggc tttagaacat
      301 gagtcttatg agctaggcga gcgccgcttc ctcaagatgc ttgagcgtca agcgaaagct
      361 ggtgagattg cagacaacgc agccgctaag ccgttactcg ctacgcttct ccctaagtta
      421 accacacgta tcgtcgagtg gctcgaagag tacgcatcga agaaaggccg caagcctagc
      481 gcatacgcac cgctccagtt actcaagccg gaggcctccg cgtttatcac cctgaaagtt
      541 atccttgcgt cactaaccag tacgaacatg acaaccattc aggccgctgc tggtatgctg
      601 gggaaagcca ttgaggacga ggcacgattt gggcgcatcc gtgacctaga agcgaagcac
      661 ttcaagaagc acgttgagga acagcttaac aagcgccacg ggcaagtcta caagaaagca
      721 tttatgcagg tggtcgaggc cgatatgatt ggtcgaggtc tgcttggtgg cgaggcgtgg
      781 tctagctggg ataaagaaac cacgatgcac gtagggattc gcctgattga aatgctgatt
      841 gaatccacgg gtctggtgga attacagcgc cacaacgcag gtaacgcagg ctctgaccat
      901 gaggcactgc aactggccca agagtacgtg gacgtattag cgaagcgtgc aggcgctctg
      961 gcgggtatct ctccgatgtt ccagccgtgt gtcgtaccgc cgaaaccttg ggtagcaatc
     1021 acagggggcg gctattgggc taacggtcgc agacctttgg cactcgttcg cactcactct
     1081 aagaagggct tgatgcgcta cgaagacgtt tacatgccag aagtctacaa ggctgtgaac
     1141 ctcgcgcaaa acaccgcatg gaaaatcaac aagaaagttc ttgctgttgt caatgagatt
     1201 gttaactgga agaattgccc ggtagcagac attccatcgc tggagcgcca agagttaccg
     1261 cctaagcctg acgacattga caccaacgag gcagcgctca aggagtggaa gaaagccgct
     1321 gctggtatct atcgcttgga caaggcacga gtgtctcgcc gtatcagctt agagttcatg
     1381 ctggagcagg ccaacaagtt cgcaagtaag aaagcaatct ggttccctta caacatggac
     1441 tggcgcggtc gtgtgtacgc tgtgccgatg ttcaacccgc aaggcaacga catgacgaaa
     1501 ggtctgctga cccttgctaa aggcaagcca atcggtgagg aaggtttcta ctggctgaaa
     1561 atccacggtg cgaactgtgc gggtgttgat aaggttccat tcccggagcg catcgcgttc
     1621 attgagaagc acgtagacga cattctggct tgcgctaaag acccaatcaa taacacttgg
     1681 tgggctgagc aggattcacc gttctgtttc ctcgcgtttt gcttcgagta tgcaggcgtt
     1741 acgcaccacg gtctgagcta caattgctct ctgccgctgg cgttcgacgg gtcttgctct
     1801 ggtatccagc acttctccgc gatgctccgc gatgaggtag gcggtcgtgc ggttaacctg
     1861 ctgccaagcg aaaccgtgca ggacatttac ggcatcgttg cacagaaagt aaacgagatt
     1921 ctcaaacagg atgcaatcaa cggcacgcct aacgagatga ttaccgtgac cgacaaggac
     1981 accggggaaa tctcagagaa gctcaaactt ggaacctcaa cgctggcgca acagtggctg
     2041 gcatatggtg taacccgtag cgtaactaaa cgttcggtca tgacgctggc ttacggttcc
     2101 aaggagttcg gctttcgtca acaggtattg gatgacacca ttcagcctgc aattgacagc
     2161 ggtaagggct tgatgttcac ccaaccgaac caagcggctg gctatatggc taagctgatt
     2221 tgggatgcgg taagcgtgac cgtagttgca gcggttgagg cgatgaactg gctcaaatct
     2281 gccgctaagc tgctggctgc tgaggtcaag gacaagaaga ccaaggagat tctgcgccac
     2341 cgttgcgcgg ttcactggac tacgccggac ggcttcccgg tctggcagga ataccgcaag
     2401 ccactccaga agcgtctcga tatgattttc ttagggcaat tccgtctgca accgacgatt
     2461 aataccctca aggattcagg cattgacgca cacaagcagg agtctggcat cgctcctaac
     2521 tttgttcact cacaggacgg tagccacctc cgcatgacag tcgtttatgc tcacgagaag
     2581 tatggcattg agtcctttgc gctcatccat gacagctttg ggactatccc ggcagacgct
     2641 ggtaagctct ttaaggctgt gcgtgaaacg atggttatca cctatgagaa caacgatgtg
     2701 ctggcagact tctactctca gtttgccgac cagctacacg agacccaact ggacaagatg
     2761 cctccgcttc cgaagaaagg aaacctgaac ctgcaagaca ttctcaagtc tgactttgcc
     2821 tttgcataat aacgactcag gctgctactc aaaactagca taaccccttg gggcctctaa
     2881 acgggtcttg aggggttttt tgggcttgat atcgaattcc tgcagcccgg ggcgcaacgc
     2941 aattaatgta agttagctca ctcattaggc accgacgtcg gaattgccag ctggggcgcc
     3001 ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat
     3061 ctgatggcgc aggggatcat gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3121 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3181 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3241 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3301 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3361 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3421 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3481 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3541 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3601 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3661 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3721 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3781 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3841 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3901 gttcttctga gcgggactct ggggttcgag agctcgcttg gactcctgtt gatagatcca
     3961 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     4021 tttattggtg agaatccaag cactagggac agtaagacgg gtaagcctgt tgatgatacc
     4081 gctgccttac tgggtgcatt agccagtctg aatgacctgt cacgggataa tccgaagtgg
     4141 tcagactgga aaatcagagg gcaggaactg ctgaacagca aaaagtcaga tagcaccaca
     4201 tagcagaccc gccataaaac gccctgagaa gcccgtgacg ggcttttctt gtattatggg
     4261 tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc catttacccc cattcactgc
     4321 cagagccgtg agcgcagcga actgaatgtc acgaaaaaga cagcgactca ggtgcctgat
     4381 ggtcggagac aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg ctacttaagc
     4441 ctttagggtt ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac atccttttgt
     4501 aatactgcgg aactgactaa agtagtgagt tatacacagg gctgggatct attcttttta
     4561 tcttttttta ttctttcttt attctataaa ttataaccac ttgaatataa acaaaaaaaa
     4621 cacacaaagg tctagcggaa tttacagagg gtctagcaga atttacaagt tttccagcaa
     4681 aggtctagca gaatttacag atacccagat cacccgggaa aaggactagt aattatcatt
     4741 gactagccca tctcaattgg tatagtgatt aaaatcacct agaccaattg agatgtatgt
     4801 ctgaattagt tgttttcaaa gcaaatgaac tagcgattag tcgctatgac ttaacggagc
     4861 atgaaaccaa gctaatttta tgctgtgtgg cactactcaa ccccacgatt gaaaacccta
     4921 caaggaaaga acggacggta tcgttcactt ataaccaata cgctcagatg atgaacatca
     4981 gtagggaaaa tgcttatggt gtattagcta aagcaaccag agagctgatg acgagaactg
     5041 tggaaatcag gaatcctttg gttaaaggct ttgagatttt ccagtggaca aactatgcca
     5101 agttctcaag cgaaaaatta gaattagttt ttagtgaaga gatattgcct tatcttttcc
     5161 agttaaaaaa attcataaaa tataatctgg aacatgttaa gtcttttgaa aacaaatact
     5221 ctatgaggat ttatgagtgg ttattaaaag aactaacaca aaagaaaact cacaaggcaa
     5281 atatagagat tagccttgat gaatttaagt tcatgttaat gcttgaaaat aactaccatg
     5341 agtttaaaag gcttaaccaa tgggttttga aaccaataag taaagattta aacacttaca
     5401 gcaatatgaa attggtggtt gataagcgag gccgcccgac tgatacgttg attttccaag
     5461 ttgaactaga tagacaaatg gatctcgtaa ccgaacttga gaacaaccag ataaaaatga
     5521 atggtgacaa aataccaaca accattacat cagattccta cctacataac ggactaagaa
     5581 aaacactaca cgatgcttta actgcaaaaa ttcagctcac cagttttgag gcaaaatttt
     5641 tgagtgacat gcaaagtaag tatgatctca atggttcgtt ctcatggctc acgcaaaaac
     5701 aacgaaccac actagagaac atactggcta aatacggaag gatctgaggt tcttatggct
     5761 cttgtatcta tcagtgaagc atcaagacta acaaacaaaa gtagaacaac tgttcaccgt
     5821 tacatatcaa agggaaaact gtccatatgc acagatgaaa acggtgtaaa aaagatagat
     5881 acatcagagc ttttacgagt ttttggtgca ttcaaagctg ttcaccatga acagatcgac
     5941 aatgtaacag atgaacagca tgtaacacct aatagaacag gtgaaaccag taaaacaaag
     6001 caactagaac atgaaattga acacctgaga caacttgtta cagctcaaca gtcacacata
     6061 gacagcctga aacaggcgat gctgcttatc gaatcaaagc tgccgacaac acgggagcca
     6121 gtgacgcctc ccgtggggaa aaaatcatgg caattctgga agaaatagcg ctttcagccg
     6181 gcaaaccggc tgaagccgga tctgcgattc tgataacaaa ctagcaacac cagaacagcc
     6241 cgtttgcggg cagcaaaacc cgtaccctag gtataaacgc agaaaggccc acccgaaggt
     6301 gagccagtgt gactctagta gagagcgttc accgacaaac aacagataaa acgaaaggcc
     6361 cagtctttcg actgagcctt tcgttttatt tgatgcctgg agatccttac tcgagtttgg
     6421 atccttaggt ctcacga
//
