LOCUS       Exported                5960 bp DNA     circular SYN 07-MAY-2025
DEFINITION  Phagemid vector pMID21, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    pMID21 vector (V004730)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5960)
  AUTHORS   Hoet RM, Coehn EH, Kent RB, Rookey K, Schoonbroodt S, Hogan S, Rem 
            L, Frans N, Daukandt M, Pieters H, van Hegelsom R, Coolen-van Neer 
            N, Nastri HG, Rondon IJ, Leeds J, Hufton SE, Huang L, Kashin I, 
            Devlin M, Kuang G, Steukers M, Viswanathan M, Nixon AE, Sexton DJ, 
            Hoogenboom HR, Ladner RC.
  TITLE     Generation of high-affinity human antibodies by combining 
            donor-derived and synthetic complementarity-determining-region 
            diversity
  JOURNAL   Nat. Biotechnol. 23 (3), 344-348 (2005)
  PUBMED    15723048
REFERENCE   2  (bases 1 to 5960)
  AUTHORS   Ladner RC, Hoogenboom HR, Hoet RM, Cohen EH, Kashin I, Rondon IJ, 
            Rem L, Frans N, Schoonbroodt S, Kent RB, Rookey K, Hogan S.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-SEP-2004) Research, Dyax Corp, 300 Technology Square, 
            Cambridge, MA 02139, USA
REFERENCE   3  (bases 1 to 5960)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5960)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 5960)
  TITLE     Direct Submission
REFERENCE   6  (bases 1 to 5960)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat.
            Biotechnol."; date: "2005"; volume: "23"; issue: "3"; pages: 
            "344-348"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-SEP-2004) Research, Dyax Corp, 300 Technology Square, Cambridge,
            MA 02139, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     SGRef: number: 5; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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)."
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2269..2325
                     /codon_start=1
                     /note="encodes light chain signal sequence; antibody
                     stuffer"
                     /translation="VKKLLFAIPLVVPFYSHSA"
     misc_feature    2326..2352
                     /label=encodes light chain antibody stuffer
                     /note="encodes light chain antibody stuffer"
     CDS             2365..2682
                     /label=hIg-kappa-CL
                     /note="Human immunoglobulin kappa light chain constant
                     region"
     CDS             2726..2788
                     /codon_start=1
                     /label=heavy chain signal
                     /note="encodes heavy chain signal sequence; antibody
                     stuffer"
                     /translation="MKKLLFMIPLVVPFVAQPASA"
     misc_feature    2867..3766
                     /label=encodes heavy chain antibody stuffer
                     /note="encodes heavy chain antibody stuffer"
     CDS             4516..4533
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             4543..4572
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
     CDS             4612..5070
                     /codon_start=1
                     /label=M13 gene III
                     /translation="SGDFDYEKMANANKGAMTENADENALQSDAKGKLDSVATDYGAAI
                     DGFIGDVSGLANGNGATGDFAGSNSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFV
                     FGAGKPYEFSIDCDKINLFRGVFAFLLYVATFMYVFSTFANILRNKES"
     primer_bind     complement(5092..5108)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(5304..5759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt tggagccttt tttttggaga ttttcaacgt gaaaaaatta
     2281 ttattcgcaa ttcctttagt tgttcctttc tattctcaca gtgcacaggt ccaactgcag
     2341 gtcgacctcg agatcaaacg tggaactgtg gctgcaccat ctgtcttcat cttcccgcca
     2401 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat
     2461 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag
     2521 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg
     2581 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc
     2641 ctgagttcac cggtgacaaa gagcttcaac aggggagagt gttaataagg cgcgcctaac
     2701 catctatttc aaggagacag tcataatgaa aaagctttta ttcatgatcc cgttagttgt
     2761 accgttcgtg gcccagccgg cctctgctga agttcaattg ttagagtctg gtggcggtct
     2821 tgttcagcct ggtggttctt tacgtctttc ttgcgctgct tccggagctt cagatctgtt
     2881 tgcctttttg tggggtggtg cagatcgcgt tacggagatc gaccgactgc ttgagcaaaa
     2941 gccacgctta actgctgatc aggcatggga tgttattcgc caaaccagtc gtcaggatct
     3001 taacctgagg ctttttttac ctactctgca agcagcgaca tctggtttga cacagagcga
     3061 tccgcgtcgt cagttggtag aaacattaac acgttgggat ggcatcaatt tgcttaatga
     3121 tgatggtaaa acctggcagc agccaggctc tgccatcctg aacgtttggc tgaccagtat
     3181 gttgaagcgt accgtagtgg ctgccgtacc tatgccattt gataagtggt acagcgccag
     3241 tggctacgaa acaacccagg acggcccaac tggttcgctg aatataagtg ttggagcaaa
     3301 aattttgtat gaggcggtgc agggagacaa atcaccaatc ccacaggcgg ttgatctgtt
     3361 tgctgggaaa ccacagcagg aggttgtgtt ggctgcgctg gaagatacct gggagactct
     3421 ttccaaacgc tatggcaata atgtgagtaa ctggaaaaca cctgcaatgg ccttaacgtt
     3481 ccgggcaaat aatttctttg gtgtaccgca ggccgcagcg gaagaaacgc gtcatcaggc
     3541 ggagtatcaa aaccgtggaa cagaaaacga tatgattgtt ttctcaccaa cgacaagcga
     3601 tcgtcctgtg cttgcctggg atgtggtcgc acccggtcag agtgggttta ttgctcccga
     3661 tggaacagtt gataagcact atgaagatca gctgaaaatg tacgaaaatt ttggccgtaa
     3721 gtcgctctgg ttaacgaagc aggatgtgga ggcgcataag gagtcgtcta gagacaactc
     3781 taagaatact ctctacttgc agatgaacag cttaagtctg agcattcggt ccgggcaaca
     3841 ttctccaaac tgaccagacg acacaaacgg cttacgctaa atcccgcgca tgggatggta
     3901 aagaggtggc gtctttgctg gcctggactc atcagatgaa ggccaaaaat tggcaggagt
     3961 ggacacagca ggcagcgaaa caagcactga ccatcaactg gtactatgct gatgtaaacg
     4021 gcaatattgg ttatgttcat actggtgctt atccagatcg tcaatcaggc catgatccgc
     4081 gattacccgt tcctggtacg ggaaaatggg actggaaagg gctattgcct tttgaaatga
     4141 accctaaggt gtataacccc cagaagctag cctgcggctt cggtcaccgt ctcaagcgcc
     4201 tccaccaagg gcccatcggt cttcccgcta gcaccctcct ccaagagcac ctctgggggc
     4261 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg
     4321 aactcaggcg ccctgaccag cggcgtccac accttcccgg ctgtcctaca gtcctcagga
     4381 ctctactccc tcagcagcgt agtgaccgtg ccctccagca gcttgggcac ccagacctac
     4441 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa
     4501 tcttgtgcgg ccgcacatca tcatcaccat cacggggccg cagaacaaaa actcatctca
     4561 gaagaggatc tgaatggggc cgcagaggct agttctgcta gtaacgcgtc ttccggtgat
     4621 tttgattatg aaaagatggc aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa
     4681 aacgcgctac agtctgacgc taaaggcaaa cttgattctg tcgctactga ttacggtgct
     4741 gctatcgatg gtttcattgg tgacgtttcc ggccttgcta atggtaatgg tgctactggt
     4801 gattttgctg gctctaattc ccaaatggct caagtcggtg acggtgataa ttcaccttta
     4861 atgaataatt tccgtcaata tttaccttcc ctccctcaat cggttgaatg tcgccctttt
     4921 gtctttggcg ctggtaaacc atatgaattt tctattgatt gtgacaaaat aaacttattc
     4981 cgtggtgtct ttgcgtttct tttatatgtt gccaccttta tgtatgtatt ttctacgttt
     5041 gctaacatac tgcgtaataa ggagtcttaa tgaaacgcgt gatgagaatt cactggccgt
     5101 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
     5161 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5221 acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc ttacgcatct
     5281 gtgcggtatt tcacaccgca tataaattgt aaacgttaat attttgttaa aattcgcgtt
     5341 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5401 taaatcaaaa gaatagcccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5461 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     5521 cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     5581 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     5641 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     5701 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgta
     5761 ctatggttgc tttgacgggt gcagtctcag tacaatctgc tctgatgccg catagttaag
     5821 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5881 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5941 gtcatcaccg aaacgcgcga
//
