LOCUS       sequence-210096-        4630 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSC101 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_42
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        18..52
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_RNA        60..109
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(152..154,155..868)
                     /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             152..865
                     /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(182..210)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     805..825
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      885..956
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      972..999
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1018..1038
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1039..1096
                     /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(1120..1137)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1671..1893
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1941..2891
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(3505..4296,4297..4365))
                     /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     4128..4147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4366..4470)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4538..4556
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4560..4603)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4606..4627
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggtgtcttcg gcgcgccttt acggctagct cagtcctagg tactatgcta gcaaggtagc
       61 tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc ctctacaaat
      121 aattttgttt aatcacacag gaaagtacta gatgagcaaa ggagaagaac ttttcactgg
      181 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtccg
      241 tggagagggt gaaggtgatg ctacaaacgg aaaactcacc cttaaattta tttgcactac
      301 tggaaaacta cctgttccgt ggccaacact tgtcactact ctgacctatg gtgttcaatg
      361 cttttcccgt tatccggatc acatgaaacg gcatgacttt ttcaagagtg ccatgcccga
      421 aggttatgta caggaacgca ctatatcttt caaagatgac gggacctaca agacgcgtgc
      481 tgaagtcaag tttgaaggtg atacccttgt taatcgtatc gagttaaagg gtattgattt
      541 taaagaagat ggaaacattc ttggacacaa actcgagtac aactttaact cacacaatgt
      601 atacatcacg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa
      661 cgttgaagat ggttccgttc aactagcaga ccattatcaa caaaatactc caattggcga
      721 tggccctgtc cttttaccag acaaccatta cctgtcgaca caatctgtcc tttcgaaaga
      781 tcccaacgaa aagcgtgacc acatggtcct tcttgagttt gtaactgctg ctgggattac
      841 acatggcatg gatgagctct acaaataata ctagagccag gcatcaaata aaacgaaagg
      901 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctctact
      961 agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttatagaaga catcgcttac
     1021 tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc
     1081 tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg
     1141 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tgacagtaag
     1201 acgggtaagc ctgttgatga taccgctgcc ttactgggtg cattagccag tctgaatgac
     1261 ctgtcacggg ataatccgaa gtggtcagac tggaaaatca gagggcagga actgctgaac
     1321 agcaaaaagt cagatagcac cacatagcag acccgccata aaacgccctg agaagcccgt
     1381 gacgggcttt tcttgtatta tgggtagttt ccttgcatga atccataaaa ggcgcctgta
     1441 gtgccattta cccccattca ctgccagagc cgtgagcgca gcgaactgaa tgtcacgaaa
     1501 aagacagcga ctcaggtgcc tgatggtcgg agacaaaagg aatattcagc gatttgcccg
     1561 agcttgcgag ggtgctactt aagcctttag ggttttaagg tctgttttgt agaggagcaa
     1621 acagcgtttg cgacatcctt ttgtaatact gcggaactga ctaaagtagt gagttataca
     1681 cagggctggg atctattctt tttatctttt tttattcttt ctttattcta taaattataa
     1741 ccacttgaat ataaacaaaa aaaacacaca aaggtctagc ggaatttaca gagggtctag
     1801 cagaatttac aagttttcca gcaaaggtct agcagaattt acagataccc acaactcaaa
     1861 ggaaaaggac tagtaattat cattgactag cccatctcaa ttggtatagt gattaaaatc
     1921 acctagacca attgagatgt atgtctgaat tagttgtttt caaagcaaat gaactagcga
     1981 ttagtcgcta tgacttaacg gagcatgaaa ccaagctaat tttatgctgt gtggcactac
     2041 tcaaccccac gattgaaaac cctacaagga aagaacggac ggtatcgttc acttataacc
     2101 aatacgctca gatgatgaac atcagtaggg aaaatgctta tggtgtatta gctaaagcaa
     2161 ccagagagct gatgacgaga actgtggaaa tcaggaatcc tttggttaaa ggctttgaga
     2221 ttttccagtg gacaaactat gccaagttct caagcgaaaa attagaatta gtttttagtg
     2281 aagagatatt gccttatctt ttccagttaa aaaaattcat aaaatataat ctggaacatg
     2341 ttaagtcttt tgaaaacaaa tactctatga ggatttatga gtggttatta aaagaactaa
     2401 cacaaaagaa aactcacaag gcaaatatag agattagcct tgatgaattt aagttcatgt
     2461 taatgcttga aaataactac catgagttta aaaggcttaa ccaatgggtt ttgaaaccaa
     2521 taagtaaaga tttaaacact tacagcaata tgaaattggt ggttgataag cgaggccgcc
     2581 cgactgatac gttgattttc caagttgaac tagatagaca aatggatctc gtaaccgaac
     2641 ttgagaacaa ccagataaaa atgaatggtg acaaaatacc aacaaccatt acatcagatt
     2701 cctacctaca taacggacta agaaaaacac tacacgatgc tttaactgca aaaattcagc
     2761 tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag taagtatgat ctcaatggtt
     2821 cgttctcatg gctcacgcaa aaacaacgaa ccacactaga gaacatactg gctaaatacg
     2881 gaaggatctg aggttcttat ggctcttgta tctatcagtg aagcatcaag actaacaaac
     2941 aaaagtagaa caactgttca ccgttacata tcaaagggaa aactgtccat atgcacagat
     3001 gaaaacggtg taaaaaagat agatacatca gagcttttac gagtttttgg tgcattcaaa
     3061 gctgttcacc atgaacagat cgacaatgta acagatgaac agcatgtaac acctaataga
     3121 acaggtgaaa ccagtaaaac aaagcaacta gaacatgaaa ttgaacacct gagacaactt
     3181 gttacagctc aacagtcaca catagacagc ctgaaacagg cgatgctgct tatcgaatca
     3241 aagctgccga caacacggga gccagtgacg cctcccgtgg ggaaaaaatc atggcaattc
     3301 tggaagaaat agcgctttca gccggcaaac cggctgaagc cggatctgcg attctgataa
     3361 caaactagca acaccagaac agcccgtttg cgggcagcaa aacccgtacc gattatcaaa
     3421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagaac cacgctcacc
     3661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4201 aaggatctta ccgctgttga gatccagttc gatataaccc actcgtgcac ccaactgatc
     4261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4501 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa
     4561 tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct
     4621 tctagaggga
//
