LOCUS       sequence-242421-        4563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For cloning and subsequent expression of peptide fragments fused 
            with Avi-tag (for biotinylation) and with pIII protein of M13K07 
            bacteriophage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4563)
  AUTHORS   Lomakin YA, Kaminskaya AN, Stepanov AV, Shmidt AA, Gabibov A, 
            Belogurov AA
  TITLE     Probing surface membrane receptors using engineered bacteriophage 
            bioconjugates.
  JOURNAL   Bioconjug Chem. 2019 Apr 25. doi: 10.1021/acs.bioconjchem.9b00218.
  PUBMED    31021608
REFERENCE   2  (bases 1 to 4563)
  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: "Bioconjug 
            Chem. 2019 Apr 25. doi: 10.1021/acs.bioconjchem.9b00218."
COMMENT     Sequence Label: pHen2 Avi-tag-linker-p3
FEATURES             Location/Qualifiers
     source          1..4563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             59..103
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1361..1378)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1361..1377)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1370..1392)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1590..2045
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1677..1696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1887..1908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2062..2081)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2181..2203
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2241..2259)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2327..2431
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2432..2500,2501..3292)
                     /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(2650..2669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3463..4051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3952..3971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4205..4222
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4339..4360
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4375..4380,4381..4398,4399..4405)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4413..4429
                     /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     4418..4440
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4437..4453
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4437..4453
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     sig_peptide     join(4505..4563,1..7)
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
ORIGIN
        1 catggcctta gctagcagtg gtggaggcgg ttcaggcgga ggtggctctg gcggtagtgg
       61 tctgaacgac atcttcgagg ctcagaaaat cgaatggcac gaaagtggtg gaggcggttc
      121 aactgttgaa agttgtttag caaaacctca tacagaaaat tcatttacta acgtctggaa
      181 agacgacaaa actttagatc gttacgctaa ctatgagggc tgtctgtgga atgctacagg
      241 cgttgtggtt tgtactggtg acgaaactca gtgttacggt acatgggttc ctattgggct
      301 tgctatccct gaaaatgagg gtggtggctc tgagggtggc ggttctgagg gtggcggttc
      361 tgagggtggc ggtactaaac ctcctgagta cggtgataca cctattccgg gctatactta
      421 tatcaaccct ctcgacggca cttatccgcc tggtactgag caaaaccccg ctaatcctaa
      481 tccttctctt gaggagtctc agcctcttaa tactttcatg tttcagaata ataggttccg
      541 aaataggcag ggtgcattaa ctgtttatac gggcactgtt actcaaggca ctgaccccgt
      601 taaaacttat taccagtaca ctcctgtatc atcaaaagcc atgtatgacg cttactggaa
      661 cggtaaattc agagactgcg ctttccattc tggctttaat gaggatccat tcgtttgtga
      721 atatcaaggc caatcgtctg acctgcctca acctcctgtc aatgctggcg gcggctctgg
      781 tggtggttct ggtggcggct ctgagggtgg cggctctgag ggtggcggct ctgagggtgg
      841 cggttctgag ggtggcggct ctgagggtgg cggttccggt ggcggctccg gttccggtga
      901 ttttgattat gaaaaaatgg caaacgctaa taagggggct atgaccgaaa atgccgatga
      961 aaacgcgcta cagtctgacg ctaaaggcaa acttgattct gtcgctactg attacggtgc
     1021 tgctatcgat ggtttcattg gtgacgtttc cggccttgct aatggtaatg gtgctactgg
     1081 tgattttgct ggctctaatt cccaaatggc tcaagtcggt gacggtgata attcaccttt
     1141 aatgaataat ttccgtcaat atttaccttc tttgcctcag tcggttgaat gtcgccctta
     1201 tgtctttggc gctggtaaac catatgaatt ttctattgat tgtgacaaaa taaacttatt
     1261 ccgtggtgtc tttgcgtttc ttttatatgt tgccaccttt atgtatgtat tttcgacgtt
     1321 tgctaacata ctgcgtaata aggagtctta ataagaattc actggccgtc gttttacaac
     1381 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1441 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     1501 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     1561 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
     1621 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc
     1681 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1741 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1801 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1861 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     1921 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt
     1981 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     2041 aattttatgg tgcagtctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
     2101 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     2161 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     2221 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2281 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2341 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2401 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2461 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2521 agtaaaagat gctgaagatc agttgggtgc ccgagtgggt tacatcgaac tggatctcaa
     2581 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2641 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2701 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2761 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2821 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2881 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2941 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3001 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3061 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3121 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3181 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3241 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3301 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3361 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3601 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3781 aacggggggt tcgtgcatac agcccagctt ggagcgaacg acctacaccg aactgagata
     3841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4081 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4141 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4201 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     4261 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     4321 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     4381 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     4441 aaacagctat gaccatgatt acgccaagct tgcatgcaaa ttctatttca aggagacagt
     4501 cataatgaaa tacctattgc ctacggcagc cgctggattg ttattactcg cggcccagcc
     4561 ggc
//
