LOCUS       Exported                4992 bp DNA     circular SYN 21-JUL-2025
DEFINITION  3rd generation plasmid for phage display on modified geneIII, 
            contains stuffer fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4992)
  AUTHORS   Andris-Widhopf J, Rader C, Steinberger P, Fuller R, Barbas CF 3rd
  TITLE     Methods for the generation of chicken monoclonal antibody fragments 
            by phage display.
  JOURNAL   J Immunol Methods. 2000 Aug 28;242(1-2):159-81.
  PUBMED    10986398
REFERENCE   2  (bases 1 to 4992)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4992)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4992)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 4992)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Immunol
            Methods."; date: "2000-08-28"; volume: "242(1-2)"; pages: "159-81"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     71..90
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     324..341
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    458..479
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        494..524
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    532..548
                     /label=lac operator
                     /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)."
     CDS             571..633
                     /codon_start=1
                     /label=OmpA signal peptide
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             complement(643..708)
                     /codon_start=1
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     primer_bind     complement(756..772)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(780..796)
                     /label=lac operator
                     /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(804..834)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(849..870)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(924..941)
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             complement(942..1253)
                     /codon_start=1
                     /label=hIg-CH
                     /translation="KGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPATVSWNSGALTS
                     GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTS
                     "
     CDS             1280..1498
                     /codon_start=1
                     /label=VL
                     /translation="WYQQKPGQAPRLLIYGTSSRATGIPDRFSGSGSGTDFTLTISRLE
                     PEDFAVYYCQQYGGSPWFGQGTKVELKR"
     CDS             1499..1816
                     /codon_start=1
                     /label=hIg-kappa-CL
                     /note="Human immunoglobulin kappa light chain constant
                     region"
                     /translation="TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA
                     LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSLPVTKSFNRG
                     EC"
     CDS             1850..1915
                     /codon_start=1
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             1934..2260
                     /codon_start=1
                     /label=TrxA
                     /note="E. coli thioredoxin"
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             2312..2329
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             2336..2362
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             2369..2902
                     /codon_start=1
                     /product="M13 pIII
                     "
                     /label=pIII
                     /translation="EGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKMANANKGAMTEN
                     ADENVLQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGSNSQMAQVGDGD
                     NSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVFAFLLYVATFMY
                     VFSTFANILRNKES"
     rep_origin      complement(2956..3411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3438..3542
                     /label=AmpR promoter
     CDS             3543..4400
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4574..4992
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
       61 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
      121 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
      181 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
      241 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
      301 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
      361 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      421 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
      481 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
      541 gataacaatt gaattcagga ggaatttaaa atgaaaaaga cagctatcgc gattgcagtg
      601 gcactggctg gtttcgctac cgtggcccag gcggccgagc tcggccatgg ctggttgggc
      661 agcgagtaat aacaatccag cggctgccgt aggcaatagg tatttcatta tgactgtctc
      721 cttggcgact agctagttta gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      781 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
      841 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
      901 tcgggaaacc tgtcgtgtta ctaatgatgg tgatggtgat ggctagtttt gtcacaagat
      961 ttgggctcaa ctttcttgtc caccttggtg ttgctgggct tgtgattcac gttgcagatg
     1021 taggtctggg tgcccaagct gctggagggc acggtcacca cgctgctgag ggagtagagt
     1081 cctgaggact gtaggacagc cgggaaggtg tgcacgccgc tggtcagggc gcctgagttc
     1141 cacgacaccg tcgccggttc ggggaagtag tccttgacca ggcagcccag ggccgctgtg
     1201 cccccagagg tgctcttgga ggagggtgcc agggggaaga ccgatgggcc cttggtggag
     1261 gctgcggaga cggtgaccgt ggtaccagca gaaacctggc caggctccca ggctcctcat
     1321 ctatggtaca tccagcaggg ccactggcat cccagacagg ttcagtggca gtgggtctgg
     1381 gacagacttc actctcacca tcagcagact ggagcctgaa gattttgcag tgtactactg
     1441 tcagcagtat ggtggctcac cgtggttcgg ccaagggacc aaggtggaac tcaaacgaac
     1501 tgtggctgca ccatctgtct tcatcttccc gccatctgat gagcagttga aatctggaac
     1561 tgcctctgtt gtgtgcctgc tgaataactt ctatcccaga gaggccaaag tacagtggaa
     1621 ggtggataac gccctccaat cgggtaactc ccaggagagt gtcacagagc aggacagcaa
     1681 ggacagcacc tacagcctca gcagcaccct gacgctgagc aaagcagact acgagaaaca
     1741 caaagtctac gcctgcgaag tcacccatca gggcctgagc ttgcccgtca caaagagctt
     1801 caacagggga gagtgttagt tctagataat taattaggag gaatttaaaa tgaaatacct
     1861 attgcctacg gcagccgctg gattgttatt actcgctgcc caaccagcca tggccgaggt
     1921 gcagctgctc gagatgagcg ataaaattat tcacctgact gacgacagtt ttgacacgga
     1981 tgtactcaaa gcggacgggg cgatcctcgt cgatttctgg gcagagtggt gcggtccgtg
     2041 caaaatgatc gccccgattc tggatgaaat cgctgacgaa tatcagggca aactgaccgt
     2101 tgcaaaactg aacatcgatc aaaaccctgg cactgcgccg aaatatggca tccgtggtat
     2161 cccgactctg ctgctgttca aaaacggtga agtggcggca accaaagtgg gtgcactgtc
     2221 taaaggtcag ttgaaagagt tcctcgacgc taacctggcg tacccgtacg acgttccgga
     2281 ctacggttct actagtggcc aggccggcca gcaccatcac catcaccatg gcgcataccc
     2341 gtacgacgtt ccggactacg cttcttagga gggtggtggc tctgagggtg gcggttctga
     2401 gggtggcggc tctgagggag gcggttccgg tggtggctct ggttccggtg attttgatta
     2461 tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa aatgccgatg aaaacgtgct
     2521 acagtctgac gctaaaggca aacttgattc tgtcgctact gattacggtg ctgctatcga
     2581 tggtttcatt ggtgacgttt ccggccttgc taatggtaat ggtgctactg gtgattttgc
     2641 tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat aattcacctt taatgaataa
     2701 tttccgtcaa tatttacctt ccctccctca atcggttgaa tgtcgccctt ttgtctttgg
     2761 cgctggtaaa ccatatgaat tttctattga ttgtgacaaa ataaacttat tccgtggtgt
     2821 ctttgcgttt cttttatatg ttgccacctt tatgtatgta ttttctacgt ttgctaacat
     2881 actgcgtaat aaggagtctt aagctagcta attaatttaa gcggccgcag atctgctctc
     2941 tgaggaggat ctgggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3001 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3061 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3121 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3181 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3241 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3301 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3361 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3421 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3481 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3541 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     3601 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     3661 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     3721 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     3781 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     3841 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3901 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3961 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4021 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4081 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4141 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     4201 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     4261 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     4321 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     4381 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     4441 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     4501 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4561 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4621 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     4681 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     4741 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     4801 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     4861 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     4921 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     4981 cgcttcccga ag
//
