LOCUS       sequence-209379-        4630 bp    DNA     circular SYN 14-APR-2025
DEFINITION  pSC101 var7 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 Apr 14, 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_49
FEATURES             Location/Qualifiers
     source          1..4630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        17..51
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     misc_RNA        59..108
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     CDS             join(151..153,154..867)
                     /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             151..864
                     /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(181..209)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     804..824
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      884..955
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      971..998
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1017..1037
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1038..1095
                     /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(1119..1136)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      1670..1892
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1940..2890
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTDYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLGNNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             complement(join(3504..4295,4296..4364))
                     /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     4127..4146
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4365..4469)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4537..4555
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(4559..4602)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4605..4626
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gtgtcttcgg cgcgccttta cggctagctc agtcctaggt actatgctag caaggtagct
       61 gtcaccggat gtgctttccg gtctgatgag tccgtgagga cgaaacagcc tctacaaata
      121 attttgttta atcacacagg aaagtactag atgagcaaag gagaagaact tttcactgga
      181 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtccgt
      241 ggagagggtg aaggtgatgc tacaaacgga aaactcaccc ttaaatttat ttgcactact
      301 ggaaaactac ctgttccgtg gccaacactt gtcactactc tgacctatgg tgttcaatgc
      361 ttttcccgtt atccggatca catgaaacgg catgactttt tcaagagtgc catgcccgaa
      421 ggttatgtac aggaacgcac tatatctttc aaagatgacg ggacctacaa gacgcgtgct
      481 gaagtcaagt ttgaaggtga tacccttgtt aatcgtatcg agttaaaggg tattgatttt
      541 aaagaagatg gaaacattct tggacacaaa ctcgagtaca actttaactc acacaatgta
      601 tacatcacgg cagacaaaca aaagaatgga atcaaagcta acttcaaaat tcgccacaac
      661 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
      721 ggccctgtcc ttttaccaga caaccattac ctgtcgacac aatctgtcct ttcgaaagat
      781 cccaacgaaa agcgtgacca catggtcctt cttgagtttg taactgctgc tgggattaca
      841 catggcatgg atgagctcta caaataatac tagagccagg catcaaataa aacgaaaggc
      901 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
      961 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatagaagac atcgcttact
     1021 agtagcggcc gctgcagtcc ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct
     1081 ttaaaaccga aaagattact tcgcgttatg caggcttcct cgctcactga ctcgctgcgc
     1141 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat gacagtaaga
     1201 cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt ctgaatgacc
     1261 tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa ctgctgaaca
     1321 gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga gaagcccgtg
     1381 acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag gcgcctgtag
     1441 tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa
     1501 agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg atttgcccga
     1561 gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta gaggagcaaa
     1621 cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg agttatacac
     1681 agggctggga tctattcttt ttatcttttt ttattctttc tttattctat aaattataac
     1741 cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag agggtctagc
     1801 agaatttaca agttttccag caaaggtcta gcagaattta cagataccca caactcaaag
     1861 gaaaaggact agtaattatc attgactagc ccatctcaat tggtatagtg attaaaatca
     1921 cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg aactagcgat
     1981 tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg tggcactact
     2041 caaccccacg attgaaaacc ctacaaggaa agaacggacg gtatcgttca cttataacca
     2101 atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag ctaaagcaac
     2161 cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag gcttcgagat
     2221 tttccagtgg acagactatg ccaagttctc aagcgaaaaa ttagaattag tttttagtga
     2281 agagatattg ccctatcttt tccagttaaa aaaattcata aaatataatc tggaacatgt
     2341 taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa aagaactaac
     2401 acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta agttcatgtt
     2461 aatgcttgga aataactacc atgagtttaa aaggcttaac caatgggttt tgaaaccaat
     2521 aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc gaggccgccc
     2581 gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg taaccgaact
     2641 tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta catcagattc
     2701 ctacctacat aacggactaa gaaaaacact acacgatgct ttaactgcaa aaattcagct
     2761 caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagtatgatc tcaatggttc
     2821 gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg ctaaatacgg
     2881 aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga ctaacaaaca
     2941 aaagtagaac aactgttcac cgttacatat caaagggaaa actgtccata tgcacagatg
     3001 aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt gcattcaaag
     3061 ctgttcacca tgaacagatc gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa
     3121 caggtgaaac cagtaaaaca aagcaactag aacatgaaat tgaacacctg agacaacttg
     3181 ttacagctca acagtcacac atagacagcc tgaaacaggc gatgctgctt atcgaatcaa
     3241 agctgccgac aacacgggag ccagtgacgc ctcccgtggg gaaaaaatca tggcaattct
     3301 ggaagaaata gcgctttcag ccggcaaacc ggctgaagcc ggatctgcga ttctgataac
     3361 aaactagcaa caccagaaca gcccgtttgc gggcagcaaa acccgtaccg attatcaaaa
     3421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg
     3661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4201 aggatcttac cgctgttgag atccagttcg atataaccca ctcgtgcacc caactgatct
     4261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4441 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4501 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
     4561 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt
     4621 ctagagggag
//
