LOCUS       sequence-205997-        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        21..55
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_RNA        63..112
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(155..157,158..871)
                     /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             155..868
                     /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(185..213)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     808..828
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      888..959
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      975..1002
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1021..1041
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1042..1099
                     /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(1123..1140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1674..1896
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1944..2894
                     /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(3508..4299,4300..4368))
                     /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     4131..4150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4369..4473)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4541..4559
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4563..4606)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4609..4630
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggaggtgtct tcggcgcgcc tttacggcta gctcagtcct aggtactatg ctagcaaggt
       61 agctgtcacc ggatgtgctt tccggtctga tgagtccgtg aggacgaaac agcctctaca
      121 aataattttg tttaatcaca caggaaagta ctagatgagc aaaggagaag aacttttcac
      181 tggagttgtc ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt
      241 ccgtggagag ggtgaaggtg atgctacaaa cggaaaactc acccttaaat ttatttgcac
      301 tactggaaaa ctacctgttc cgtggccaac acttgtcact actctgacct atggtgttca
      361 atgcttttcc cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc
      421 cgaaggttat gtacaggaac gcactatatc tttcaaagat gacgggacct acaagacgcg
      481 tgctgaagtc aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga
      541 ttttaaagaa gatggaaaca ttcttggaca caaactcgag tacaacttta actcacacaa
      601 tgtatacatc acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca
      661 caacgttgaa gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg
      721 cgatggccct gtccttttac cagacaacca ttacctgtcg acacaatctg tcctttcgaa
      781 agatcccaac gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat
      841 tacacatggc atggatgagc tctacaaata atactagagc caggcatcaa ataaaacgaa
      901 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
      961 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttataga agacatcgct
     1021 tactagtagc ggccgctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt
     1081 ttctttaaaa ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct
     1141 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatgacagt
     1201 aagacgggta agcctgttga tgataccgct gccttactgg gtgcattagc cagtctgaat
     1261 gacctgtcac gggataatcc gaagtggtca gactggaaaa tcagagggca ggaactgctg
     1321 aacagcaaaa agtcagatag caccacatag cagacccgcc ataaaacgcc ctgagaagcc
     1381 cgtgacgggc ttttcttgta ttatgggtag tttccttgca tgaatccata aaaggcgcct
     1441 gtagtgccat ttacccccat tcactgccag agccgtgagc gcagcgaact gaatgtcacg
     1501 aaaaagacag cgactcaggt gcctgatggt cggagacaaa aggaatattc agcgatttgc
     1561 ccgagcttgc gagggtgcta cttaagcctt tagggtttta aggtctgttt tgtagaggag
     1621 caaacagcgt ttgcgacatc cttttgtaat actgcggaac tgactaaagt agtgagttat
     1681 acacagggct gggatctatt ctttttatct ttttttattc tttctttatt ctataaatta
     1741 taaccacttg aatataaaca aaaaaaacac acaaaggtct agcggaattt acagagggtc
     1801 tagcagaatt tacaagtttt ccagcaaagg tctagcagaa tttacagata cccacaactc
     1861 aaaggaaaag gactagtaat tatcattgac tagcccatct caattggtat agtgattaaa
     1921 atcacctaga ccaattgaga tgtatgtctg aattagttgt tttcaaagca aatgaactag
     1981 cgattagtcg ctatgactta acggagcatg aaaccaagct aattttatgc tgtgtggcac
     2041 tactcaaccc cacgattgaa aaccctacaa ggaaagaacg gacggtatcg ttcacttata
     2101 accaatacgc tcagatgatg aacatcagta gggaaaatgc ttatggtgta ttagctaaag
     2161 caaccagaga gctgatgacg agaactgtgg aaatcaggaa tcctttggtt aaaggctttg
     2221 ggattttcca gtggacaaac tatgccaagt tctcaagcga aaaattagaa ttagttttta
     2281 gtgaagagat attgccttat cttttccagt taaaaaaatt cataaaatat aatctggaac
     2341 atgttaagtc ttttgaaaac aaatactcta tgaggattta tgagtggtta ttaaaagaac
     2401 taacacaaaa gaaaactcac aaggcaaata tagagattag ccttgatgaa tttaagttca
     2461 tgttaatgct tgaaaataac taccatgagt ttaaaaggct taaccaatgg gttttgaaac
     2521 caataagtaa agatttaaac acttacagca atatgaaatt ggtggttgat aagcgaggcc
     2581 gcccgactga tacgttgatt ttccaagttg aactagatag acaaatggat ctcgtaaccg
     2641 aacttgagaa caaccagata aaaatgaatg gtgacaaaat accaacaacc attacatcag
     2701 attcctacct acataacgga ctaagaaaaa cactacacga tgctttaact gcaaaaattc
     2761 agctcaccag ttttgaggca aaatttttga gtgacatgca aagtaagtat gatctcaatg
     2821 gttcgttctc atggctcacg caaaaacaac gaaccacact agagaacata ctggctaaat
     2881 acggaaggat ctgaggttct tatggctctt gtatctatca gtgaagcatc aagactaaca
     2941 aacaaaagta gaacaactgt tcaccgttac atatcaaagg gaaaactgtc catatgcaca
     3001 gatgaaaacg gtgtaaaaaa gatagataca tcagagcttt tacgagtttt tggtgcattc
     3061 aaagctgttc accatgaaca gatcgacaat gtaacagatg aacagcatgt aacacctaat
     3121 agaacaggtg aaaccagtaa aacaaagcaa ctagaacatg aaattgaaca cctgagacaa
     3181 cttgttacag ctcaacagtc acacatagac agcctgaaac aggcgatgct gcttatcgaa
     3241 tcaaagctgc cgacaacacg ggagccagtg acgcctcccg tggggaaaaa atcatggcaa
     3301 ttctggaaga aatagcgctt tcagccggca aaccggctga agccggatct gcgattctga
     3361 taacaaacta gcaacaccag aacagcccgt ttgcgggcag caaaacccgt accgattatc
     3421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     3481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag aaccacgctc
     3661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     4021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     4081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     4141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     4201 ctcaaggatc ttaccgctgt tgagatccag ttcgatataa cccactcgtg cacccaactg
     4261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     4321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     4381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     4441 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     4501 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggca
     4561 gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc
     4621 gcttctagag
//
