LOCUS       pComb3X        3451 bp DNA     circular SYN 20-JUL-2025
DEFINITION  Phagemid cloning vector pComb3X, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3451)
  AUTHORS   Rader C, Barbas CF III.
  JOURNAL   (in) PHAGE DISPLAY, A LABORATORY MANUAL. Cold Spring Harbor 
            Laboratory Press, Cold Spring Harbor, NY, USA (2000), In press
REFERENCE   2  (bases 1 to 3451)
  AUTHORS   Rader C, Barbas CF III.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-MAY-2000) Department of Molecular Biology, BCC-526, 
            Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 
            92037, USA
REFERENCE   3  (bases 1 to 3451)
  AUTHORS   Rader C, Fuller R, Barbas CF III.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-SEP-2003) Department of Molecular Biology, BCC-526, 
            Scripps Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 
            92037, USA
REFERENCE   4  (bases 1 to 3451)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3451)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 3451)
  TITLE     Direct Submission
REFERENCE   7  (bases 1 to 3451)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "(in) PHAGE
            DISPLAY, A LABORATORY MANUAL. Cold Spring Harbor Laboratory Press,
            Cold Spring Harbor, NY, USA (2000), In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (16-MAY-2000) Department of Molecular Biology, BCC-526, Scripps
            Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037,
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted
            (05-SEP-2003) Department of Molecular Biology, BCC-526, Scripps
            Research Institute, 10550 N. Torrey Pines Rd., La Jolla, CA 92037,
            USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
COMMENT     SGRef: number: 6; type: "Journal Article"
COMMENT     On Sep 5, 2003 this sequence version replaced AF268281.1.
FEATURES             Location/Qualifiers
     source          1..3451
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    109..130
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        145..175
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    183..199
                     /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)."
     sig_peptide     222..284
                     /label=OmpA signal peptide
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
     sig_peptide     329..394
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     CDS             440..457
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             464..490
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             497..1030
                     /codon_start=1
                     /label=M13 PIII
                     /translation="EGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKMANANKGAMTEN
                     ADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGSNSQMAQVGDGD
                     NSPLMNNFRQYLPSLPQSVECRPFVFSAGKPYEFSIDCDKINLFRGVFAFLLYVATFMY
                     VFSTFANILRNKES"
     rep_origin      complement(1066..1521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1548..1652
                     /label=AmpR promoter
     CDS             1653..2510
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2684..3272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 ctcatgcatt
      301 ctagataatt aattaggagg aatttaaaat gaaataccta ttgcctacgg cagccgctgg
      361 attgttatta ctcgctgccc aaccagccat ggccgaggtg cagctgctcg agggatccac
      421 tagtggccag gccggccagc accatcacca tcaccatggc gcatacccgt acgacgttcc
      481 ggactacgct tcttaggagg gtggtggctc tgagggtggc ggttctgagg gtggcggctc
      541 tgagggaggc ggttccggtg gtggctctgg ttccggtgat tttgattatg aaaagatggc
      601 aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc
      661 taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg gtttcattgg
      721 tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg gctctaattc
      781 ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt tccgtcaata
      841 tttaccttcc ctccctcaat cggttgaatg tcgccctttt gtctttagcg ctggtaaacc
      901 atatgaattt tctattgatt gtgacaaaat aaacttattc cgtggtgtct ttgcgtttct
      961 tttatatgtt gccaccttta tgtatgtatt ttctacgttt gctaacatac tgcgtaataa
     1021 ggagtcttaa gctagctaat taatttaagc ggccgcagat ctgggaaatt gtaagcgtta
     1081 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1141 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1201 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1261 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1321 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1381 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1441 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1501 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1561 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1621 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1681 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1741 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1801 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1861 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1921 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1981 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2041 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2101 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2161 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2221 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2281 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2341 ctgataaatc tggagccggt gagcgtggat ctcgcggtat cattgcagca ctggggccag
     2401 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2461 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2521 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2581 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2641 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2701 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2761 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2821 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2881 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2941 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3001 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3061 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3121 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3181 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3241 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3301 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3361 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3421 agcgcagcga gtcagtgagc gaggaagcgg a
//
