LOCUS       sequence-462690-        4467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Periplasmic Expresssion of Strep-tagged anti-Zika NbD32.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4467)
  AUTHORS   Yuli Peng, Atheer Alqatari, Fabian Kiessling, Dominik Renn, Raik 
            Grunberg, Stefan T. Arold,, Magnus Rueping
  TITLE     Nanobody-Based Lateral Flow Assay for Rapid Zika Virus Detection
  JOURNAL   ACS Synthetic Biology Article ASAP
REFERENCE   2  (bases 1 to 4467)
  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"; journalName: "ACS 
            Synthetic Biology Article ASAP"
COMMENT     Sequence Label: pJEx411c_pelB-N32-Strep2
FEATURES             Location/Qualifiers
     source          1..4467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             415..438
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             451..474
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     terminator      492..539
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(762..781)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     promoter        848..939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             940..1749
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1832..2914
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(1851..1870)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      3000..3588
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3489..3508
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3742..3759
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3857..3874
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3858..3874
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4338..4357
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4338..4356
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4361..4377
                     /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)."
     RBS             4450..4458
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     sig_peptide     join(4465..4467,1..63)
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
ORIGIN
        1 aaatacctgc tgccgaccgc tgctgctggt ctgctgctcc tcgctgccca gccggcgatg
       61 gccgaagttc agctcgtcga aagcggcggt ggtctggttc aagcgggtgg ctctttgcgt
      121 ctgtcgtgcg cggtatctgg aattgacttc tcccgctatg ctattacgtg gaaccgccag
      181 tcgccgggga accagcgccg agagtgggtg gccacattac cgccggccga taccactgtt
      241 tatgcagatg ctgtcaaagg gcgttttacc atctcacggg acaatacgaa gaataccgtg
      301 taccttcaga tgaacagcct gaaacctgaa gatactgcgg tgtactattg tgcaaccagc
      361 ccacgtatcc ataattgggg ccaaggcacg caggtaaccg tgagtagtag cggcttggaa
      421 gtactgttcc agggtccaac cggctctgca tggagccacc cgcaatttga gaaagtagta
      481 aactagtccc cctagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      541 ccctgagagg cgtcaatcga gttcgtacct aagggcgaca ccccctaatt agcccgggcg
      601 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
      661 tcgcatgggg agtccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
      721 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa acaaggggtg ttatgagcca
      781 tattcaggta taaatgggct cgcgataatg ttcagaattg gttaattggt tgtaacactg
      841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      901 ccctgataaa tgcttcaata atattgaaaa aggaagaata tgagccatat tcaacgggaa
      961 acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1021 cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg
     1081 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1141 gtcagactaa actggctgac ggaatttatg ccacttccga ccatcaagca ttttatccgt
     1201 actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc gttccaggta
     1261 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     1321 cggttgcact cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc
     1381 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     1441 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     1501 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     1561 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     1621 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     1681 aaatatggta ttgataatcc tgatatgaat aaattgcaat ttcatttgat gctcgatgag
     1741 tttttctaag cggcgcgcca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg
     1801 cccggaagag agtcaattca gggtggtgaa tatgaaacca gtaacgttat acgatgtcgc
     1861 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     1921 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     1981 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     2041 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     2101 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     2161 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     2221 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     2281 ctctgaccag acacccatca acagtattat tttctcccat gaggacggta cgcgactggg
     2341 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     2401 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     2461 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     2521 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     2581 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     2641 agtgggatac gacgataccg aagatagctc atgttatatc ccgccgttaa ccaccatcaa
     2701 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     2761 ccaggcggtg aagggcaatc agctgttgcc agtctcactg gtgaaaagaa aaaccaccct
     2821 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     2881 acgacaggtt tcccgactgg aaagcgggca gtgactcatg accaaaatcc cttaacgtga
     2941 gttacgcgcg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3001 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3061 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3121 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagc ccaccacttc
     3181 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3241 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3301 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3361 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3421 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     3481 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     3541 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     3601 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     3661 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     3721 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaagg cgagagtagg
     3781 gaactgccag gcatcaaact aagcagaagg cccctgacgg atggcctttt tgcgtttcta
     3841 caaactcttt ctgtgttgta aaacgacggc cagtcttaag ctcgggcccc ctgggcggtt
     3901 ctgataacga gtaatcgtta atccgcaaat aacgtaaaaa cccgcttcgg cgggtttttt
     3961 tatgggggga gtttagggaa agagcatttg tcagaatatt taagggcgcc tgtcactttg
     4021 cttgatatat gagaattatt taaccttata aatgagaaaa aagcaacgca ctttaaataa
     4081 gatacgttgc tttttcgatt gatgaacacc tataattaaa ctattcatct attatttatg
     4141 attttttgta tatacaatat ttctagtttg ttaaagagaa ttaagaaaat aaatctcgaa
     4201 aataataaag ggaaaatcag tttttgatat caaaattata catgtcaacg ataatacaaa
     4261 atataataca aactataaga tgttatcagt atttattatg catttagaat aaattttgtg
     4321 tcgcccttcc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg ataacaattg
     4381 ggcccaagtt cacttaaaaa ggagatcaac aatgaaagca attttcgtac tgaaacatct
     4441 taatcatgct aaggaggttt tctaatg
//
