LOCUS       sequence-202871-        3923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3923)
  AUTHORS   Lim KH, Huang H, Pralle A, Park S
  TITLE     Stable, high-affinity streptavidin monomer for protein labeling and 
            monovalent biotin detection.
  JOURNAL   Biotechnol Bioeng. 2013 Jan;110(1):57-67. doi: 10.1002/bit.24605. 
            Epub 2012 Aug 8.
  PUBMED    22806584
REFERENCE   2  (bases 1 to 3923)
  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"; doi: "10.1002/bit.24605";
            journalName: "Biotechnol Bioeng"; date: "2013-01"; volume: "110"; 
            issue: "1"; pages: "57-67"
COMMENT     Sequence Label: pRSET-mSA
FEATURES             Location/Qualifiers
     source          1..3923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     21..40
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             71..93
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             148..489
                     /codon_start=1
                     /product="monomeric streptavidin/rhizavidin hybrid (Lim et 
                     al., 2013)"
                     /label=mSA
                     /translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
                     TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
                     QGQDTFTKVK"
     CDS             511..534
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             553..1266
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLFKDPNEKRDHMVLLE
                     FLTAAGITEGMNELYK"
     primer_bind     complement(1325..1343)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1339..1386
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1483..1938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1570..1589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1780..1801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1964..2068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2069..2137,2138..2929)
                     /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     complement(2287..2306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3100..3688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3589..3608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3842..3859
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ggatctcgat cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc
       61 tagaaataat tttgtttaac tttaagaagg agatatacca tgggcagcag ccatcaccat
      121 catcaccaca gccaggatct ggctagcgcg gaagcgggta tcaccggcac gtggtacaac
      181 cagtctggtt ctaccttcac cgttaccgcg ggtgcggacg gtaacctgac cggtcagtac
      241 gaaaaccgtg cgcagggcac tggttgccag aactctccgt acaccctgac cggtcgttac
      301 aacggtacca aactggaatg gcgtgttgaa tggaacaact ctaccgaaaa ctgccactct
      361 cgtaccgaat ggcgtggtca gtaccagggt ggtgcggaag cgcgtatcaa cacccagtgg
      421 aacctgacct acgaaggtgg ttctggtccg gcgaccgaac agggtcagga caccttcacc
      481 aaagttaaac cgtctgcggc gtctggatcc gactacaagg acgatgacga caagtaactc
      541 gagggcggat ctatgtctaa aggtgaagaa ttattcactg gtgttgtccc aattttggtt
      601 gaattagatg gtgatgttaa tggtcacaaa ttttctgtct ccggtgaagg tgaaggtgat
      661 gctacgtacg gtaaattgac cttaaaatta ctctgtacta ctggtaaatt gccagttcca
      721 tggccaacct tagtcactac tttaggttat ggtgttcaat gttttgctag atacccagat
      781 catatgaaac aacatgactt tttcaagtct gccatgccag aaggttatgt tcaagaaaga
      841 actatttttt tcaaagatga cggtaactac aagaccagag ctgaagtcaa gtttgaaggt
      901 gataccttag ttaatagaat cgaattaaaa ggtattgatt ttaaagaaga tggtaacatt
      961 ttaggtcaca aattggaata caactataac tctcacaatg tttacatcac tgctgacaaa
     1021 caaaagaatg gtatcaaagc taacttcaaa attagacaca acattgaaga tggtggtgtt
     1081 caattagctg accattatca acaaaatact ccaattggtg atggtccagt cttgttacca
     1141 gacaaccatt acttatccta tcaatctaag ttattcaaag atccaaacga aaagagagac
     1201 cacatggtct tgttagaatt tttgactgct gctggtatta ccgagggtat gaatgaattg
     1261 tacaaataaa agcttgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     1321 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1381 tttttgctga aaggaggaac tatatccgga tctggcgtaa tagcgaagag gcccgcaccg
     1441 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     1501 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1561 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1621 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1681 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1741 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     1801 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     1861 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     1921 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1981 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2041 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2101 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2161 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2221 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2281 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2341 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2401 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2461 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2521 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2581 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2641 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2701 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2761 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2821 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2881 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     2941 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3001 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3061 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3121 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3181 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3241 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3301 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3361 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3421 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3481 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3541 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3601 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3661 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3721 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3781 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3841 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     3901 gcgttggccg attcattaat gca
//
