LOCUS       sequence-156504-        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-EGFP
FEATURES             Location/Qualifiers
     source          1..3923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     75..94
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        75..93
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             125..147
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             166..183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             202..543
                     /codon_start=1
                     /product="monomeric streptavidin/rhizavidin hybrid (Lim et 
                     al., 2013)"
                     /label=mSA
                     /translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
                     TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
                     QGQDTFTKVK"
     CDS             565..588
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(604..606,607..609,610..1323)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(604..606,607..609,610..1320)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(649..670)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(910..929)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1257..1278
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1379..1397)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1393..1440
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1537..1992
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1624..1643)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1834..1855
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2018..2122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2123..2191,2192..2983)
                     /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(2341..2360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3154..3742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3643..3662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3896..3913
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcaggatct
       61 cgatcccgcg aaattaatac gactcactat agggagacca caacggtttc cctctagaaa
      121 taattttgtt taactttaag aaggagatat accatgggca gcagccatca ccatcatcac
      181 cacagccagg atctggctag cgcggaagcg ggtatcaccg gcacgtggta caaccagtct
      241 ggttctacct tcaccgttac cgcgggtgcg gacggtaacc tgaccggtca gtacgaaaac
      301 cgtgcgcagg gcactggttg ccagaactct ccgtacaccc tgaccggtcg ttacaacggt
      361 accaaactgg aatggcgtgt tgaatggaac aactctaccg aaaactgcca ctctcgtacc
      421 gaatggcgtg gtcagtacca gggtggtgcg gaagcgcgta tcaacaccca gtggaacctg
      481 acctacgaag gtggttctgg tccggcgacc gaacagggtc aggacacctt caccaaagtt
      541 aaaccgtctg cggcgtctgg atccgactac aaggacgatg acgacaagct cgagggcggt
      601 agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      661 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      721 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      781 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      841 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      901 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      961 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1021 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1081 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1141 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1201 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1261 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1321 taaaagcttg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     1381 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1441 ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc accgatcgcc
     1501 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     1561 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1621 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1681 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1741 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1801 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1861 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1921 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1981 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2041 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2101 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2161 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2221 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2281 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     2341 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2401 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2461 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2521 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2581 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2641 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2701 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2761 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2821 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2881 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2941 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3001 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3061 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     3241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     3301 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     3361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     3421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     3481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     3541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     3601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     3661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     3721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     3781 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     3841 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     3901 gtcagtgagc gaggaagcgg aag
//
