LOCUS       sequence-257663-        6490 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 6490)
  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 6490)
  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: pYD1-mSA
FEATURES             Location/Qualifiers
     source          1..6490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             138..149
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             150..176
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             240..581
                     /codon_start=1
                     /product="monomeric streptavidin/rhizavidin hybrid (Lim et 
                     al., 2013)"
                     /label=mSA
                     /translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
                     TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
                     QGQDTFTKVK"
     CDS             603..626
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(956..975)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(957..975)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(982..999)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(982..998)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(991..1013)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1139..1594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1226..1245)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1436..1457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1697..2371)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2372..2652)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(2673..2692)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2792..2814
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2852..2870)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    2911..3414
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3451..3555
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3556..3624,3625..4416)
                     /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(3774..3793)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4587..5175
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5076..5095
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5329..5346
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5463..5484
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5499..5504,5505..5522,5523..5529)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5537..5553
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     5542..5564
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5561..5577
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5561..5577
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5598..5616
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5646..6310
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    5862..5979
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     6274..6297
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 acgggaaggc aatgcaagga gtttttgaat attacaaatc agtaacgttt gtcagtaatt
       61 gcggttctca cccctcaaca actagcaaag gcagccccat aaacacacag tatgttttta
      121 aggacaatag ctcgacgatt gaaggtagat acccatacga cgttccagac tacgctctgc
      181 aggctagtgg tggtggtggt tctggtggtg gtggttctgg tggtggtggt tctgctagcg
      241 cggaagcggg tatcaccggc acgtggtaca accagtctgg ttctaccttc accgttaccg
      301 cgggtgcgga cggtaacctg accggtcagt acgaaaaccg tgcgcagggc actggttgcc
      361 agaactctcc gtacaccctg accggtcgtt acaacggtac caaactggaa tggcgtgttg
      421 aatggaacaa ctctaccgaa aactgccact ctcgtaccga atggcgtggt cagtaccagg
      481 gtggtgcgga agcgcgtatc aacacccagt ggaacctgac ctacgaaggt ggttctggtc
      541 cggcgaccga acagggtcag gacaccttca ccaaagttaa accgtctgcg gcgtctggat
      601 ccgactacaa ggacgatgac gacaagtaac tcgagatctg ataacaacag tgtagatgta
      661 acaaaatcga ctttgttccc actgtacttt tagctcgtac aaaatacaat atacttttca
      721 tttctccgta aacaacatgt tttcccatgt aatatccttt tctatttttc gttccgttac
      781 caactttaca catactttat atagctattc acttctatac actaaaaaac taagacaatt
      841 ttaattttgc tgcctgccat atttcaattt gttataaatt cctataattt atcctattag
      901 tagctaaaaa aagatgaatg tgaatcgaat cctaagagaa ttgagctcca attcgcccta
      961 tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     1021 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     1081 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggac
     1141 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1201 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1261 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1321 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1381 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1441 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1501 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1561 gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     1621 atctgtgcgg tatttcacac cgcatagatc ggcaagtgca caaacaatac ttaaataaat
     1681 actactcagt aataacctat ttcttagcat ttttgacgaa atttgctatt ttgttagagt
     1741 cttttacacc atttgtctcc acacctccgc ttacatcaac accaataacg ccatttaatc
     1801 taagcgcatc accaacattt tctggcgtca gtccaccagc taacataaaa tgtaagcttt
     1861 cggggctctc ttgccttcca acccagtcag aaatcgagtt ccaatccaaa agttcacctg
     1921 tcccacctgc ttctgaatca aacaagggaa taaacgaatg aggtttctgt gaagctgcac
     1981 tgagtagtat gttgcagtct tttggaaata cgagtctttt aataactggc aaaccgagga
     2041 actcttggta ttcttgccac gactcatctc catgcagttg gacgatatca atgccgtaat
     2101 cattgaccag agccaaaaca tcctccttag gttgattacg aaacacgcca accaagtatt
     2161 tcggagtgcc tgaactattt ttatatgctt ttacaagact tgaaattttc cttgcaataa
     2221 ccgggtcaat tgttctcttt ctattgggca cacatataat acccagcaag tcagcatcgg
     2281 aatctagagc acattctgcg gcctctgtgc tctgcaagcc gcaaactttc accaatggac
     2341 cagaactacc tgtgaaatta ataacagaca tactccaagc tgcctttgtg tgcttaatca
     2401 cgtatactca cgtgctcaat agtcaccaat gccctccctc ttggccctct ccttttcttt
     2461 tttcgaccga attaattctt aatcggcaaa aaaagaaaag ctccggatca agattgtacg
     2521 taaggtgaca agctattttt caataaagaa tatcttccac tactgccatc tggcgtcata
     2581 actgcaaagt acacatatat tacgatgctg ttctattaaa tgcttcctat attatatata
     2641 tagtaatgtc gtgatctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2701 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2761 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     2821 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2881 aatgtcatga taataatggt ttcttagacg gatcgcttgc ctgtaactta cacgcgcctc
     2941 gtatctttta atgatggaat aatttgggaa tttactctgt gtttatttat ttttatgttt
     3001 tgtatttgga ttttagaaag taaataaaga aggtagaaga gttacggaat gaagaaaaaa
     3061 aaataaacaa aggtttaaaa aatttcaaca aaaagcgtac tttacatata tatttattag
     3121 acaagaaaag cagattaaat agatatacat tcgattaacg ataagtaaaa tgtaaaatca
     3181 caggattttc gtgtgtggtc ttctacacag acaagatgaa acaattcggc attaatacct
     3241 gagagcagga agagcaagat aaaaggtagt atttgttggc gatcccccta gagtctttta
     3301 catcttcgga aaacaaaaac tattttttct ttaatttctt tttttacttt ctatttttaa
     3361 tttatatatt tatattaaaa aatttaaatt ataattattt ttatagcacg tgatgaaaag
     3421 gacccaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3481 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3541 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3601 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3661 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3721 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3781 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3841 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3901 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3961 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4021 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4081 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4141 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4201 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4261 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4321 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4381 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4441 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4501 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4561 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4621 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4681 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4741 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4801 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4861 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4921 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4981 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5041 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5101 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5161 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5221 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5281 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5341 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5401 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5461 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     5521 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     5581 gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctggt accaattcct
     5641 tgaattttca aaaattctta cttttttttt ggatggacgc aaagaagttt aataatcata
     5701 ttacatggca ttaccaccat atacatatcc atatacatat ccatatctaa tcttacttat
     5761 atgttgtgga aatgtaaaga gccccattat cttagcctaa aaaaaccttc tctttggaac
     5821 tttcagtaat acgcttaact gctcattgct atattgaagt acggattaga agccgccgag
     5881 cgggtgacag ccctccgaag gaagactctc ctccgtgcgt cctcgtcttc accggtcgcg
     5941 ttcctgaaac gcagatgtgc ctcgcgccgc actgctccga acaataaaga ttctacaata
     6001 ctagctttta tggttatgaa gaggaaaaat tggcagtaac ctggccccac aaaccttcaa
     6061 atgaacgaat caaattaaca accataggat gataatgcga ttagtttttt agccttattt
     6121 ctggggtaat taatcagcga agcgatgatt tttgatctat taacagatat ataaatgcaa
     6181 aaactgcata accactttaa ctaatacttt caacattttc ggtttgtatt acttcttatt
     6241 caaatgtaat aaaagtatca acaaaaaatt gttaatatac ctctatactt taacgtcaag
     6301 gagaaaaacc ccggatcgaa ttccctactt catacatttt caattaagat gcagttactt
     6361 cgctgttttt caatattttc tgttattgct tcagttttag cacaggaact gacaactata
     6421 tgcgagcaaa tcccctcacc aactttagaa tcgacgccgt actctttgtc aacgactact
     6481 attttggcca
//
