LOCUS       sequence-119143-        4934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  2nd 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 4934)
  AUTHORS   Barbas CF, Wagner J
  TITLE     Synthetic Human Antibodies: Selecting and Evolving Functional 
            Proteins Methods, A companion to Methods in Enzymology 8, 94-103
  JOURNAL   Methods, A companion to Methods in Enzymology, 1995; 8(2): 94-103.
REFERENCE   2  (bases 1 to 4934)
  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: "Methods, A 
            companion to Methods in Enzymology,"; date: "1995"; volume: " 8(2)";
            pages: " 94-103"
COMMENT     Sequence Label: pComb3HSS
FEATURES             Location/Qualifiers
     source          1..4934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    109..130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(145..150,151..168,169..175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    183..199
                     /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     222..284
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     sig_peptide     complement(293..358)
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     primer_bind     complement(406..422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(406..422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(419..441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    430..446
                     /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(454..460,461..478,479..484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(574..591)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     sig_peptide     1500..1565
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             1584..1910
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     rep_origin      complement(2549..3004)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2686..2707)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2898..2917
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3031..3135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3136..3204,3205..3996)
                     /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(3354..3373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4167..4755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4656..4675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4909..4926
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
       61 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
      121 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      181 aattgtgagc ggataacaat tgaattcagg aggaatttaa aatgaaaaag acagctatcg
      241 cgattgcagt ggcactggct ggtttcgcta ccgtggccca ggcggccgag ctcgccatgg
      301 ctggttgggc agcgagtaat aacaatccag cggctgccgt aggcaatagg tatttcatta
      361 tgactgtctc cttggcgact agctagttta gaattcgtaa tcatggtcat agctgtttcc
      421 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      481 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      541 cgctttccag tcgggaaacc tgtcgtgtta ctaatgatgg tgatggtgat ggctagtttt
      601 gtcacaagat ttgggctcaa ctttcttgtc caccttggtg ttgctgggct tgtgattcac
      661 gttgcagatg taggtctggg tgcccaagct gctggagggc acggtcacca cgctgctgag
      721 ggagtagagt cctgaggact gtaggacagc cgggaaggtg tgcacgccgc tggtcagggc
      781 gcctgagttc cacgacaccg tcgccggttc ggggaagtag tccttgacca ggcagcccag
      841 ggccgctgtg cccccagagg tgctcttgga ggagggtgcc agggggaaga ccgatgggcc
      901 cttggtggag gctgcggaga cggtgaccgt ggtaccagca gaaacctggc caggctccca
      961 ggctcctcat ctatggtaca tccagcaggg ccactggcat cccagacagg ttcagtggca
     1021 gtgggtctgg gacagacttc actctcacca tcagcagact ggagcctgaa gattttgcag
     1081 tgtactactg tcagcagtat ggtggctcac cgtggttcgg ccaagggacc aaggtggaac
     1141 tcaaacgaac tgtggctgca ccatctgtct tcatcttccc gccatctgat gagcagttga
     1201 aatctggaac tgcctctgtt gtgtgcctgc tgaataactt ctatcccaga gaggccaaag
     1261 tacagtggaa ggtggataac gccctccaat cgggtaactc ccaggagagt gtcacagagc
     1321 aggacagcaa ggacagcacc tacagcctca gcagcaccct gacgctgagc aaagcagact
     1381 acgagaaaca caaagtctac gcctgcgaag tcacccatca gggcctgagc ttgcccgtca
     1441 caaagagctt caacagggga gagtgttagt tctagataat taattaggag gaatttaaaa
     1501 tgaaatacct attgcctacg gcagccgctg gattgttatt actcgctgcc caaccagcca
     1561 tggccgaggt gcagctgctc gagatgagcg ataaaattat tcacctgact gacgacagtt
     1621 ttgacacgga tgtactcaaa gcggacgggg cgatcctcgt cgatttctgg gcagagtggt
     1681 gcggtccgtg caaaatgatc gccccgattc tggatgaaat cgctgacgaa tatcagggca
     1741 aactgaccgt tgcaaaactg aacatcgatc aaaaccctgg cactgcgccg aaatatggca
     1801 tccgtggtat cccgactctg ctgctgttca aaaacggtga agtggcggca accaaagtgg
     1861 gtgcactgtc taaaggtcag ttgaaagagt tcctcgacgc taacctggcg tacccgtacg
     1921 acgttccgga ctacggttct actagtggcc aggccggcca ggagggtggt ggctctgagg
     1981 gtggcggttc tgagggtggc ggctctgagg gaggcggttc cggtggtggc tctggttccg
     2041 gtgattttga ttatgaaaag atggcaaacg ctaataaggg ggctatgacc gaaaatgccg
     2101 atgaaaacgc gctacagtct gacgctaaag gcaaacttga ttctgtcgct actgattacg
     2161 gtgctgctat cgatggtttc attggtgacg tttccggcct tgctaatggt aatggtgcta
     2221 ctggtgattt tgctggctct aattcccaaa tggctcaagt cggtgacggt gataattcac
     2281 ctttaatgaa taatttccgt caatatttac cttccctccc tcaatcggtt gaatgtcgcc
     2341 cttttgtctt tggcgctggt aaaccatatg aattttctat tgattgtgac aaaataaact
     2401 tattccgtgg tgtctttgcg tttcttttat atgttgccac ctttatgtat gtattttcta
     2461 cgtttgctaa catactgcgt aataaggagt cttaagctag ctaattaatt taagcggccg
     2521 cagatctgct ctctgaggag gatctgggaa attgtaagcg ttaatatttt gttaaaattc
     2581 gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc
     2641 ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag
     2701 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     2761 gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
     2821 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     2881 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
     2941 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     3001 gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3061 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3121 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3181 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3241 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3301 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3361 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3421 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3481 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3541 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3601 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3661 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3721 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3781 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3841 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3901 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3961 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4021 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4081 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4141 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4201 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4261 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     4321 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4381 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4441 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4501 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4561 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4621 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4681 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4741 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4801 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4861 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     4921 agcgaggaag cgga
//
