LOCUS       sequence-473419-        6435 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 6435)
  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 6435)
  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: pDisplay-mSA-EGFP-TM
FEATURES             Location/Qualifiers
     source          1..6435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        316..519
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     470..490
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     564..583
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        564..582
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     663..722
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             726..752
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             777..1118
                     /codon_start=1
                     /product="monomeric streptavidin/rhizavidin hybrid (Lim et 
                     al., 2013)"
                     /label=mSA
                     /translation="AEAGITGTWYNQSGSTFTVTAGADGNLTGQYENRAQGTGCQNSPY
                     TLTGRYNGTKLEWRVEWNNSTENCHSRTEWRGQYQGGAEARINTQWNLTYEGGSGPATE
                     QGQDTFTKVK"
     CDS             1140..1163
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1173..1175,1176..1178,1179..1889)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1173..1175,1176..1178,1179..1889)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1218..1239)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1479..1498)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1826..1847
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1911..1940
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1944..2090
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(2112..2129)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2118..2342
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2474..3062)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2554..2573)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3391..3438
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3463..3482
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3670..4464)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             complement(3670..4464)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3781..3800)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4391..4410
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(4563..4582)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        4661..4852
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4809..4829
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4912..5703,5704..5772))
                     /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     5535..5554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5773..5877)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5904..6359
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5991..6010)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6201..6222
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        join(6371..6435,1..315)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
       61 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      121 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      181 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      241 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      301 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      361 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      421 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      481 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
      541 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agcttggtac
      601 cgagctcgga tccactagta acggccgcca gtgtgctgga attcggcttg gggatatcca
      661 ccatggagac agacacactc ctgctatggg tactgctgct ctgggttcca ggttccactg
      721 gtgactatcc atatgatgtt ccagattatg ctggggccca gccggccaga tctatggcgg
      781 aagcgggtat caccggcacg tggtacaacc agtctggttc taccttcacc gttaccgcgg
      841 gtgcggacgg taacctgacc ggtcagtacg aaaaccgtgc gcagggcact ggttgccaga
      901 actctccgta caccctgacc ggtcgttaca acggtaccaa actggaatgg cgtgttgaat
      961 ggaacaactc taccgaaaac tgccactctc gtaccgaatg gcgtggtcag taccagggtg
     1021 gtgcggaagc gcgtatcaac acccagtgga acctgaccta cgaaggtggt tctggtccgg
     1081 cgaccgaaca gggtcaggac accttcacca aagttaaacc gtctgcggcg tctggatccg
     1141 actacaagga cgatgacgac aagggcgcgc cgatggtgag caagggcgag gagctgttca
     1201 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1261 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1321 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1381 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1441 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1501 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1561 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1621 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1681 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1741 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1801 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1861 tcactctcgg catggacgag ctgtacaagc ggccgcggct gcaggtcgac gaacaaaaac
     1921 tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc atcgtggtgc
     1981 cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg gtggtgctca
     2041 ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt taggcggccg
     2101 ctcgagatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2161 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     2221 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     2281 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     2341 ggcttctgag gcggaaagaa ccagtggcgg taatacggtt atccacagaa tcaggggata
     2401 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2461 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2521 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2581 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2641 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2701 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2761 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2821 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2881 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2941 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     3001 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     3061 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     3121 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     3181 aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg gcagggcctg
     3241 ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg
     3301 aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt
     3361 gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg tgcgttttat
     3421 tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt
     3481 cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg cgctggagga
     3541 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     3601 tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt tcgaacccca
     3661 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     3721 agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc
     3781 aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca
     3841 gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc
     3901 atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg cgaacagttc
     3961 ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc
     4021 catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc
     4081 cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg
     4141 agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct
     4201 tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca
     4261 cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt cggtcttgac
     4321 aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat
     4381 tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc
     4441 gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatcgatctt
     4501 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggccg
     4561 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
     4621 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
     4681 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
     4741 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
     4801 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc aggactcttc
     4861 ctttttcaat aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4921 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4981 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5041 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5101 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5161 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5221 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5281 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5341 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5401 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5461 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5521 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5581 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5641 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5701 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5761 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5821 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5881 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5941 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     6001 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     6061 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     6121 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     6181 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     6241 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     6301 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     6361 cgcgcgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     6421 gttcatagcc catat
//
