LOCUS       sequence-210097-        4630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSC101 var2 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_44
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..58
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_RNA        66..115
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(158..160,161..874)
                     /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             158..871
                     /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(188..216)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     811..831
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      891..962
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      978..1005
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1024..1044
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1045..1102
                     /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(1126..1143)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1677..1899
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1947..2897
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFWIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(3511..4302,4303..4371))
                     /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     4134..4153
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4372..4476)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4544..4562
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4566..4609)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(4612..4630,1..3)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gagggaggtg tcttcggcgc gcctttacgg ctagctcagt cctaggtact atgctagcaa
       61 ggtagctgtc accggatgtg ctttccggtc tgatgagtcc gtgaggacga aacagcctct
      121 acaaataatt ttgtttaatc acacaggaaa gtactagatg agcaaaggag aagaactttt
      181 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
      241 tgtccgtgga gagggtgaag gtgatgctac aaacggaaaa ctcaccctta aatttatttg
      301 cactactgga aaactacctg ttccgtggcc aacacttgtc actactctga cctatggtgt
      361 tcaatgcttt tcccgttatc cggatcacat gaaacggcat gactttttca agagtgccat
      421 gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga cctacaagac
      481 gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaagggtat
      541 tgattttaaa gaagatggaa acattcttgg acacaaactc gagtacaact ttaactcaca
      601 caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg
      661 ccacaacgtt gaagatggtt ccgttcaact agcagaccat tatcaacaaa atactccaat
      721 tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat ctgtcctttc
      781 gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg
      841 gattacacat ggcatggatg agctctacaa ataatactag agccaggcat caaataaaac
      901 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
      961 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat agaagacatc
     1021 gcttactagt agcggccgct gcagtccggc aaaaaagggc aaggtgtcac caccctgccc
     1081 tttttcttta aaaccgaaaa gattacttcg cgttatgcag gcttcctcgc tcactgactc
     1141 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatgac
     1201 agtaagacgg gtaagcctgt tgatgatacc gctgccttac tgggtgcatt agccagtctg
     1261 aatgacctgt cacgggataa tccgaagtgg tcagactgga aaatcagagg gcaggaactg
     1321 ctgaacagca aaaagtcaga tagcaccaca tagcagaccc gccataaaac gccctgagaa
     1381 gcccgtgacg ggcttttctt gtattatggg tagtttcctt gcatgaatcc ataaaaggcg
     1441 cctgtagtgc catttacccc cattcactgc cagagccgtg agcgcagcga actgaatgtc
     1501 acgaaaaaga cagcgactca ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt
     1561 tgcccgagct tgcgagggtg ctacttaagc ctttagggtt ttaaggtctg ttttgtagag
     1621 gagcaaacag cgtttgcgac atccttttgt aatactgcgg aactgactaa agtagtgagt
     1681 tatacacagg gctgggatct attcttttta tcttttttta ttctttcttt attctataaa
     1741 ttataaccac ttgaatataa acaaaaaaaa cacacaaagg tctagcggaa tttacagagg
     1801 gtctagcaga atttacaagt tttccagcaa aggtctagca gaatttacag atacccacaa
     1861 ctcaaaggaa aaggactagt aattatcatt gactagccca tctcaattgg tatagtgatt
     1921 aaaatcacct agaccaattg agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac
     1981 tagcgattag tcgctatgac ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg
     2041 cactactcaa ccccacgatt gaaaacccta caaggaaaga acggacggta tcgttcactt
     2101 ataaccaata cgctcagatg atgaacatca gtagggaaaa tgcttatggt gtattagcta
     2161 aagcaaccag agagctgatg acgagaactg tggaaatcag gaatcctttg gttaaaggct
     2221 tttggatttt ccagtggaca aactatgcca agttctcaag cgaaaaatta gaattagttt
     2281 ttagtgaaga gatattgcct tatcttttcc agttaaaaaa attcataaaa tataatctgg
     2341 aacatgttaa gtcttttgaa aacaaatact ctatgaggat ttatgagtgg ttattaaaag
     2401 aactaacaca aaagaaaact cacaaggcaa atatagagat tagccttgat gaatttaagt
     2461 tcatgttaat gcttgaaaat aactaccatg agtttaaaag gcttaaccaa tgggttttga
     2521 aaccaataag taaagattta aacacttaca gcaatatgaa attggtggtt gataagcgag
     2581 gccgcccgac tgatacgttg attttccaag ttgaactaga tagacaaatg gatctcgtaa
     2641 ccgaacttga gaacaaccag ataaaaatga atggtgacaa aataccaaca accattacat
     2701 cagattccta cctacataac ggactaagaa aaacactaca cgatgcttta actgcaaaaa
     2761 ttcagctcac cagttttgag gcaaaatttt tgagtgacat gcaaagtaag tatgatctca
     2821 atggttcgtt ctcatggctc acgcaaaaac aacgaaccac actagagaac atactggcta
     2881 aatacggaag gatctgaggt tcttatggct cttgtatcta tcagtgaagc atcaagacta
     2941 acaaacaaaa gtagaacaac tgttcaccgt tacatatcaa agggaaaact gtccatatgc
     3001 acagatgaaa acggtgtaaa aaagatagat acatcagagc ttttacgagt ttttggtgca
     3061 ttcaaagctg ttcaccatga acagatcgac aatgtaacag atgaacagca tgtaacacct
     3121 aatagaacag gtgaaaccag taaaacaaag caactagaac atgaaattga acacctgaga
     3181 caacttgtta cagctcaaca gtcacacata gacagcctga aacaggcgat gctgcttatc
     3241 gaatcaaagc tgccgacaac acgggagcca gtgacgcctc ccgtggggaa aaaatcatgg
     3301 caattctgga agaaatagcg ctttcagccg gcaaaccggc tgaagccgga tctgcgattc
     3361 tgataacaaa ctagcaacac cagaacagcc cgtttgcggg cagcaaaacc cgtaccgatt
     3421 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3481 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3541 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3601 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg
     3661 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3721 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3781 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3841 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3901 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3961 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4021 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4081 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4141 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4201 actctcaagg atcttaccgc tgttgagatc cagttcgata taacccactc gtgcacccaa
     4261 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4321 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4381 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4441 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4501 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     4561 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg
     4621 gccgcttcta
//
