LOCUS       sequence-449948-        4992 bp    DNA     circular SYN 15-DEC-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)
  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: "J Immunol 
            Methods."; date: "2000-08-28"; volume: "242(1-2)"; pages: "159-81"
COMMENT     Sequence Label: pComb3XSS
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     sig_peptide     complement(3..68)
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     primer_bind     complement(116..132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(116..132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(129..151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    140..156
                     /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(164..170,171..188,189..194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    209..230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(284..301)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     sig_peptide     1210..1275
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             1294..1620
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             1672..1689
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1696..1722
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      complement(2316..2771)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2453..2474)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2665..2684
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2798..2902
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2903..2971,2972..3763)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRR
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3121..3140)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3934..4522
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4423..4442
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4676..4693
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4810..4831
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4846..4851,4852..4869,4870..4876)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4884..4900
                     /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)."
     sig_peptide     4923..4985
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
ORIGIN
        1 tcggccatgg ctggttgggc agcgagtaat aacaatccag cggctgccgt aggcaatagg
       61 tatttcatta tgactgtctc cttggcgact agctagttta gaattcgtaa tcatggtcat
      121 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
      181 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
      241 gctcactgcc cgctttccag tcgggaaacc tgtcgtgtta ctaatgatgg tgatggtgat
      301 ggctagtttt gtcacaagat ttgggctcaa ctttcttgtc caccttggtg ttgctgggct
      361 tgtgattcac gttgcagatg taggtctggg tgcccaagct gctggagggc acggtcacca
      421 cgctgctgag ggagtagagt cctgaggact gtaggacagc cgggaaggtg tgcacgccgc
      481 tggtcagggc gcctgagttc cacgacaccg tcgccggttc ggggaagtag tccttgacca
      541 ggcagcccag ggccgctgtg cccccagagg tgctcttgga ggagggtgcc agggggaaga
      601 ccgatgggcc cttggtggag gctgcggaga cggtgaccgt ggtaccagca gaaacctggc
      661 caggctccca ggctcctcat ctatggtaca tccagcaggg ccactggcat cccagacagg
      721 ttcagtggca gtgggtctgg gacagacttc actctcacca tcagcagact ggagcctgaa
      781 gattttgcag tgtactactg tcagcagtat ggtggctcac cgtggttcgg ccaagggacc
      841 aaggtggaac tcaaacgaac tgtggctgca ccatctgtct tcatcttccc gccatctgat
      901 gagcagttga aatctggaac tgcctctgtt gtgtgcctgc tgaataactt ctatcccaga
      961 gaggccaaag tacagtggaa ggtggataac gccctccaat cgggtaactc ccaggagagt
     1021 gtcacagagc aggacagcaa ggacagcacc tacagcctca gcagcaccct gacgctgagc
     1081 aaagcagact acgagaaaca caaagtctac gcctgcgaag tcacccatca gggcctgagc
     1141 ttgcccgtca caaagagctt caacagggga gagtgttagt tctagataat taattaggag
     1201 gaatttaaaa tgaaatacct attgcctacg gcagccgctg gattgttatt actcgctgcc
     1261 caaccagcca tggccgaggt gcagctgctc gagatgagcg ataaaattat tcacctgact
     1321 gacgacagtt ttgacacgga tgtactcaaa gcggacgggg cgatcctcgt cgatttctgg
     1381 gcagagtggt gcggtccgtg caaaatgatc gccccgattc tggatgaaat cgctgacgaa
     1441 tatcagggca aactgaccgt tgcaaaactg aacatcgatc aaaaccctgg cactgcgccg
     1501 aaatatggca tccgtggtat cccgactctg ctgctgttca aaaacggtga agtggcggca
     1561 accaaagtgg gtgcactgtc taaaggtcag ttgaaagagt tcctcgacgc taacctggcg
     1621 tacccgtacg acgttccgga ctacggttct actagtggcc aggccggcca gcaccatcac
     1681 catcaccatg gcgcataccc gtacgacgtt ccggactacg cttcttagga gggtggtggc
     1741 tctgagggtg gcggttctga gggtggcggc tctgagggag gcggttccgg tggtggctct
     1801 ggttccggtg attttgatta tgaaaagatg gcaaacgcta ataagggggc tatgaccgaa
     1861 aatgccgatg aaaacgtgct acagtctgac gctaaaggca aacttgattc tgtcgctact
     1921 gattacggtg ctgctatcga tggtttcatt ggtgacgttt ccggccttgc taatggtaat
     1981 ggtgctactg gtgattttgc tggctctaat tcccaaatgg ctcaagtcgg tgacggtgat
     2041 aattcacctt taatgaataa tttccgtcaa tatttacctt ccctccctca atcggttgaa
     2101 tgtcgccctt ttgtctttgg cgctggtaaa ccatatgaat tttctattga ttgtgacaaa
     2161 ataaacttat tccgtggtgt ctttgcgttt cttttatatg ttgccacctt tatgtatgta
     2221 ttttctacgt ttgctaacat actgcgtaat aaggagtctt aagctagcta attaatttaa
     2281 gcggccgcag atctgctctc tgaggaggat ctgggaaatt gtaagcgtta atattttgtt
     2341 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2401 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2461 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2521 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2581 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2641 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2701 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2761 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2821 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2881 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2941 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     3001 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     3061 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     3121 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3181 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3241 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3301 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3361 gggggatcat gtaactcgcc ttgatcgtag ggaaccggag ctgaatgaag ccataccaaa
     3421 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3481 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3541 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3601 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3661 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3721 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     3781 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     3841 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     3901 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     3961 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4021 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     4081 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     4141 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     4201 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     4261 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     4321 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     4381 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     4441 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     4501 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     4561 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     4621 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     4681 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4741 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4801 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4861 tccggctcgt atgttgtgtg gaattgtgag cggataacaa ttgaattcag gaggaattta
     4921 aaatgaaaaa gacagctatc gcgattgcag tggcactggc tggtttcgct accgtggccc
     4981 aggcggccga gc
//
