LOCUS       sequence-66341-e        3351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Template for the amplification of spiking anchors in BeSD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3351)
  AUTHORS   Diamante L, Gatti-Lafranconi P, Schaerli Y, Hollfelder F
  TITLE     In vitro affinity screening of protein and peptide binders by 
            megavalent bead surface display.
  JOURNAL   Protein Eng Des Sel. 2013 Oct;26(10):713-24. doi: 
            10.1093/protein/gzt039. Epub 2013 Aug 26.
  PUBMED    23980186
REFERENCE   2  (bases 1 to 3351)
  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: 
            "10.1093/protein/gzt039"; journalName: "Protein Eng Des Sel"; date: 
            "2013-10"; volume: "26"; issue: "10"; pages: "713-24"
COMMENT     Sequence Label: pIVEX-anchor
FEATURES             Location/Qualifiers
     source          1..3351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(33..55)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     155..174
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     521..540
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(658..676)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      672..719
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1085..1090,1091..1107,1108..1113))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1134..1150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1134..1150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1147..1169)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1158..1174
                     /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)."
     promoter        complement(join(1182..1188,1189..1206,1207..1212))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1227..1248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1365..1382)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1536..2124)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1616..1635)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2295..3086,3087..3155))
                     /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     2918..2937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3156..3260)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3328..3346
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
       61 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
      121 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
      181 gtgcaccata tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
      241 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      301 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      361 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt gcatgcaagg
      421 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
      481 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
      541 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
      601 tcgagatcga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      661 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      721 tgaaaggagg aactatatcc ggatatccac aggacgggtg tggtcgccat gatcgcgtag
      781 tcgatagtgg ctccaagtag cgaagcgagc aggactgggc ggcggccaaa gcggtcggac
      841 agtgctccga gaacgggtgc gcatagaaat tgcatcaacg catatagcgc tagcagcacg
      901 ccatagtgac tggcgatgct gtcggaatgg acgatatccc gcaagaggcc cggcagtacc
      961 ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat gacgatgagc
     1021 gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg tgataaacta
     1081 ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcgtaat catgtcatag
     1141 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1201 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1261 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1321 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1381 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1441 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1501 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1561 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1621 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1681 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1741 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1801 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1861 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1921 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     1981 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2041 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2101 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2161 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2221 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2281 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2341 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2401 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2461 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     2521 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     2581 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     2641 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     2701 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     2761 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     2821 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     2881 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     2941 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3001 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3061 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3121 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3241 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     3301 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt t
//
