LOCUS       sequence-209374-        4630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSC101 var4 BbsI cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4630)
  AUTHORS   Segall-Shapiro TH, Sontag ED, Voigt CA
  TITLE     Engineered promoters enable constant gene expression at any copy 
            number in bacteria.
  JOURNAL   Nat Biotechnol. 2018 Mar 19. pii: nbt.4111. doi: 10.1038/nbt.4111.
  PUBMED    29553576
REFERENCE   2  (bases 1 to 4630)
  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 
            Biotechnol. 2018 Mar 19. pii: nbt.4111. doi: 10.1038/nbt.4111."
COMMENT     Sequence Label: pTHSSe_46
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..53
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_RNA        61..110
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(153..155,156..869)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             153..866
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(183..211)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     806..826
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      886..957
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      973..1000
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1019..1039
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1040..1097
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1121..1138)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1672..1894
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1942..2892
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFGIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(3506..4297,4298..4366))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4129..4148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4367..4471)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4539..4557
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4561..4604)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4607..4628
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aggtgtcttc ggcgcgcctt tacggctagc tcagtcctag gtactatgct agcaaggtag
       61 ctgtcaccgg atgtgctttc cggtctgatg agtccgtgag gacgaaacag cctctacaaa
      121 taattttgtt taatcacaca ggaaagtact agatgagcaa aggagaagaa cttttcactg
      181 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtcc
      241 gtggagaggg tgaaggtgat gctacaaacg gaaaactcac ccttaaattt atttgcacta
      301 ctggaaaact acctgttccg tggccaacac ttgtcactac tctgacctat ggtgttcaat
      361 gcttttcccg ttatccggat cacatgaaac ggcatgactt tttcaagagt gccatgcccg
      421 aaggttatgt acaggaacgc actatatctt tcaaagatga cgggacctac aagacgcgtg
      481 ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaag ggtattgatt
      541 ttaaagaaga tggaaacatt cttggacaca aactcgagta caactttaac tcacacaatg
      601 tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attcgccaca
      661 acgttgaaga tggttccgtt caactagcag accattatca acaaaatact ccaattggcg
      721 atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgtc ctttcgaaag
      781 atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct gctgggatta
      841 cacatggcat ggatgagctc tacaaataat actagagcca ggcatcaaat aaaacgaaag
      901 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
      961 tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttatagaag acatcgctta
     1021 ctagtagcgg ccgctgcagt ccggcaaaaa agggcaaggt gtcaccaccc tgcccttttt
     1081 ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc ctcgctcact gactcgctgc
     1141 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atgacagtaa
     1201 gacgggtaag cctgttgatg ataccgctgc cttactgggt gcattagcca gtctgaatga
     1261 cctgtcacgg gataatccga agtggtcaga ctggaaaatc agagggcagg aactgctgaa
     1321 cagcaaaaag tcagatagca ccacatagca gacccgccat aaaacgccct gagaagcccg
     1381 tgacgggctt ttcttgtatt atgggtagtt tccttgcatg aatccataaa aggcgcctgt
     1441 agtgccattt acccccattc actgccagag ccgtgagcgc agcgaactga atgtcacgaa
     1501 aaagacagcg actcaggtgc ctgatggtcg gagacaaaag gaatattcag cgatttgccc
     1561 gagcttgcga gggtgctact taagccttta gggttttaag gtctgttttg tagaggagca
     1621 aacagcgttt gcgacatcct tttgtaatac tgcggaactg actaaagtag tgagttatac
     1681 acagggctgg gatctattct ttttatcttt ttttattctt tctttattct ataaattata
     1741 accacttgaa tataaacaaa aaaaacacac aaaggtctag cggaatttac agagggtcta
     1801 gcagaattta caagttttcc agcaaaggtc tagcagaatt tacagatacc cacaactcaa
     1861 aggaaaagga ctagtaatta tcattgacta gcccatctca attggtatag tgattaaaat
     1921 cacctagacc aattgagatg tatgtctgaa ttagttgttt tcaaagcaaa tgaactagcg
     1981 attagtcgct atgacttaac ggagcatgaa accaagctaa ttttatgctg tgtggcacta
     2041 ctcaacccca cgattgaaaa ccctacaagg aaagaacgga cggtatcgtt cacttataac
     2101 caatacgctc agatgatgaa catcagtagg gaaaatgctt atggtgtatt agctaaagca
     2161 accagagagc tgatgacgag aactgtggaa atcaggaatc ctttggttaa aggctttggg
     2221 attttccagt ggacaaacta tgccaagttc tcaagcgaaa aattagaatt agtttttagt
     2281 gaagagatat tgccttatct tttccagtta aaaaaattca taaaatataa tctggaacat
     2341 gttaagtctt ttgaaaacaa atactctatg aggatttatg agtggttatt aaaagaacta
     2401 acacaaaaga aaactcacaa ggcaaatata gagattagcc ttgatgaatt taagttcatg
     2461 ttaatgcttg aaaataacta ccatgagttt aaaaggctta accaatgggt tttgaaacca
     2521 ataagtaaag atttaaacac ttacagcaat atgaaattgg tggttgataa gcgaggccgc
     2581 ccgactgata cgttgatttt ccaagttgaa ctagatagac aaatggatct cgtaaccgaa
     2641 cttgagaaca accagataaa aatgaatggt gacaaaatac caacaaccat tacatcagat
     2701 tcctacctac ataacggact aagaaaaaca ctacacgatg ctttaactgc aaaaattcag
     2761 ctcaccagtt ttgaggcaaa atttttgagt gacatgcaaa gtaagtatga tctcaatggt
     2821 tcgttctcat ggctcacgca aaaacaacga accacactag agaacatact ggctaaatac
     2881 ggaaggatct gaggttctta tggctcttgt atctatcagt gaagcatcaa gactaacaaa
     2941 caaaagtaga acaactgttc accgttacat atcaaaggga aaactgtcca tatgcacaga
     3001 tgaaaacggt gtaaaaaaga tagatacatc agagctttta cgagtttttg gtgcattcaa
     3061 agctgttcac catgaacaga tcgacaatgt aacagatgaa cagcatgtaa cacctaatag
     3121 aacaggtgaa accagtaaaa caaagcaact agaacatgaa attgaacacc tgagacaact
     3181 tgttacagct caacagtcac acatagacag cctgaaacag gcgatgctgc ttatcgaatc
     3241 aaagctgccg acaacacggg agccagtgac gcctcccgtg gggaaaaaat catggcaatt
     3301 ctggaagaaa tagcgctttc agccggcaaa ccggctgaag ccggatctgc gattctgata
     3361 acaaactagc aacaccagaa cagcccgttt gcgggcagca aaacccgtac cgattatcaa
     3421 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3481 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3541 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3601 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac
     3661 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3721 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3781 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3841 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3901 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3961 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4021 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4081 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4141 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4201 caaggatctt accgctgttg agatccagtt cgatataacc cactcgtgca cccaactgat
     4261 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4321 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4381 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4441 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4501 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggcaga
     4561 atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat tcgcggccgc
     4621 ttctagaggg
//
