LOCUS       sequence-209377-        4630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSC101 var6 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_48
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..60
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_RNA        68..117
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(160..162,163..876)
                     /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             160..873
                     /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(190..218)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     813..833
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      893..964
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      980..1007
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1026..1046
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1047..1104
                     /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(1128..1145)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1679..1901
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1949..2899
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTDYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(3513..4304,4305..4373))
                     /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     4136..4155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4374..4478)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4546..4564
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4568..4611)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(4614..4630,1..5)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tagagggagg tgtcttcggc gcgcctttac ggctagctca gtcctaggta ctatgctagc
       61 aaggtagctg tcaccggatg tgctttccgg tctgatgagt ccgtgaggac gaaacagcct
      121 ctacaaataa ttttgtttaa tcacacagga aagtactaga tgagcaaagg agaagaactt
      181 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      241 tctgtccgtg gagagggtga aggtgatgct acaaacggaa aactcaccct taaatttatt
      301 tgcactactg gaaaactacc tgttccgtgg ccaacacttg tcactactct gacctatggt
      361 gttcaatgct tttcccgtta tccggatcac atgaaacggc atgacttttt caagagtgcc
      421 atgcccgaag gttatgtaca ggaacgcact atatctttca aagatgacgg gacctacaag
      481 acgcgtgctg aagtcaagtt tgaaggtgat acccttgtta atcgtatcga gttaaagggt
      541 attgatttta aagaagatgg aaacattctt ggacacaaac tcgagtacaa ctttaactca
      601 cacaatgtat acatcacggc agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt
      661 cgccacaacg ttgaagatgg ttccgttcaa ctagcagacc attatcaaca aaatactcca
      721 attggcgatg gccctgtcct tttaccagac aaccattacc tgtcgacaca atctgtcctt
      781 tcgaaagatc ccaacgaaaa gcgtgaccac atggtccttc ttgagtttgt aactgctgct
      841 gggattacac atggcatgga tgagctctac aaataatact agagccaggc atcaaataaa
      901 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
      961 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atagaagaca
     1021 tcgcttacta gtagcggccg ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc
     1081 cctttttctt taaaaccgaa aagattactt cgcgttatgc aggcttcctc gctcactgac
     1141 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaatg
     1201 acagtaagac gggtaagcct gttgatgata ccgctgcctt actgggtgca ttagccagtc
     1261 tgaatgacct gtcacgggat aatccgaagt ggtcagactg gaaaatcaga gggcaggaac
     1321 tgctgaacag caaaaagtca gatagcacca catagcagac ccgccataaa acgccctgag
     1381 aagcccgtga cgggcttttc ttgtattatg ggtagtttcc ttgcatgaat ccataaaagg
     1441 cgcctgtagt gccatttacc cccattcact gccagagccg tgagcgcagc gaactgaatg
     1501 tcacgaaaaa gacagcgact caggtgcctg atggtcggag acaaaaggaa tattcagcga
     1561 tttgcccgag cttgcgaggg tgctacttaa gcctttaggg ttttaaggtc tgttttgtag
     1621 aggagcaaac agcgtttgcg acatcctttt gtaatactgc ggaactgact aaagtagtga
     1681 gttatacaca gggctgggat ctattctttt tatctttttt tattctttct ttattctata
     1741 aattataacc acttgaatat aaacaaaaaa aacacacaaa ggtctagcgg aatttacaga
     1801 gggtctagca gaatttacaa gttttccagc aaaggtctag cagaatttac agatacccac
     1861 aactcaaagg aaaaggacta gtaattatca ttgactagcc catctcaatt ggtatagtga
     1921 ttaaaatcac ctagaccaat tgagatgtat gtctgaatta gttgttttca aagcaaatga
     1981 actagcgatt agtcgctatg acttaacgga gcatgaaacc aagctaattt tatgctgtgt
     2041 ggcactactc aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg tatcgttcac
     2101 ttataaccaa tacgctcaga tgatgaacat cagtagggaa aatgcttatg gtgtattagc
     2161 taaagcaacc agagagctga tgacgagaac tgtggaaatc aggaatcctt tggttaaagg
     2221 ctttgagatt ttccagtgga cagactatgc caagttctca agcgaaaaat tagaattagt
     2281 ttttagtgaa gagatattgc cttatctttt ccagttaaaa aaattcataa aatataatct
     2341 ggaacatgtt aagtcttttg aaaacaaata ctctatgagg atttatgagt ggttattaaa
     2401 agaactaaca caaaagaaaa ctcacaaggc aaatatagag attagccttg atgaatttaa
     2461 gttcatgtta atgcttgaaa ataactacca tgagtttaaa aggcttaacc aatgggtttt
     2521 gaaaccaata agtaaagatt taaacactta cagcaatatg aaattggtgg ttgataagcg
     2581 aggccgcccg actgatacgt tgattttcca agttgaacta gatagacaaa tggatctcgt
     2641 aaccgaactt gagaacaacc agataaaaat gaatggtgac aaaataccaa caaccattac
     2701 atcagattcc tacctacata acggactaag aaaaacacta cacgatgctt taactgcaaa
     2761 aattcagctc accagttttg aggcaaaatt tttgagtgac atgcaaagta agtatgatct
     2821 caatggttcg ttctcatggc tcacgcaaaa acaacgaacc acactagaga acatactggc
     2881 taaatacgga aggatctgag gttcttatgg ctcttgtatc tatcagtgaa gcatcaagac
     2941 taacaaacaa aagtagaaca actgttcacc gttacatatc aaagggaaaa ctgtccatat
     3001 gcacagatga aaacggtgta aaaaagatag atacatcaga gcttttacga gtttttggtg
     3061 cattcaaagc tgttcaccat gaacagatcg acaatgtaac agatgaacag catgtaacac
     3121 ctaatagaac aggtgaaacc agtaaaacaa agcaactaga acatgaaatt gaacacctga
     3181 gacaacttgt tacagctcaa cagtcacaca tagacagcct gaaacaggcg atgctgctta
     3241 tcgaatcaaa gctgccgaca acacgggagc cagtgacgcc tcccgtgggg aaaaaatcat
     3301 ggcaattctg gaagaaatag cgctttcagc cggcaaaccg gctgaagccg gatctgcgat
     3361 tctgataaca aactagcaac accagaacag cccgtttgcg ggcagcaaaa cccgtaccga
     3421 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3481 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3541 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3601 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
     3661 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3721 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3781 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3841 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3901 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3961 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4021 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4081 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4141 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4201 aaactctcaa ggatcttacc gctgttgaga tccagttcga tataacccac tcgtgcaccc
     4261 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4321 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4381 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4441 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4501 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     4561 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
     4621 cggccgcttc
//
