LOCUS       Exported                4766 bp DNA     circular SYN 21-JUL-2025
DEFINITION  Backbone of pComb3X containing TT Fab (light and heavy chains) 
            against tetanus toxin, used as phage display/expression control and 
            PCR template for human antibody constant regions (kappa and CH1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4766)
  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 4766)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4766)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4766)
  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"
FEATURES             Location/Qualifiers
     source          1..4766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        141..171
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     218..280
                     /label=OmpA signal peptide
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
     CDS             602..919
                     /codon_start=1
                     /label=hIg-kappa-CL
                     /note="Human immunoglobulin kappa light chain constant
                     region"
                     /translation="TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA
                     LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSLPVTKSFNRG
                     EC"
     sig_peptide     953..1018
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     CDS             1733..1750
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1757..1783
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     rep_origin      complement(2377..2832)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2859..2963
                     /label=AmpR promoter
     CDS             2964..3821
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3995..4583
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4737..4754
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaattgaa ttcaggagga atttaaaatg aaaaagacag ctatcgcgat
      241 tgcagtggca ctggctggtt tcgctaccgt ggcccaggcg gccgagctca cgcagtctcc
      301 aggcaccctg tctttgtctc caggggaaag agccaccctc tcctgcaggg ccagtcacag
      361 tgttagcagg gcctacttag cctggtacca gcagaaacct ggccaggctc ccaggctcct
      421 catctatggt acatccagca gggccactgg catcccagac aggttcagtg gcagtgggtc
      481 tgggacagac ttcactctca ccatcagcag actggagcct gaagattttg cagtgtacta
      541 ctgtcagcag tatggtggct caccgtggtt cggccaaggg accaaggtgg aactcaaacg
      601 aactgtggct gcaccatctg tcttcatctt cccgccatct gatgagcagt tgaaatctgg
      661 aactgcctct gttgtgtgcc tgctgaataa cttctatccc agagaggcca aagtacagtg
      721 gaaggtggat aacgccctcc aatcgggtaa ctcccaggag agtgtcacag agcaggacag
      781 caaggacagc acctacagcc tcagcagcac cctgacgctg agcaaagcag actacgagaa
      841 acacaaagtc tacgcctgcg aagtcaccca tcagggcctg agcttgcccg tcacaaagag
      901 cttcaacagg ggagagtgtt agttctagat aattaattag gaggaattta aaatgaaata
      961 cctattgcct acggcagccg ctggattgtt attactcgct gcccaaccag ccatggccga
     1021 ggtgcagctg ctcgagcagt ctggggctga ggtgaagaag cctgggtcct cggtgaaggt
     1081 ctcctgcagg gcttctggag gcaccttcaa caattatgcc atcagctggg tgcgacaggc
     1141 ccctggacaa gggcttgagt ggatgggagg gatcttccct ttccgtaata cagcaaagta
     1201 cgcacaacac ttccagggca gagtcaccat taccgcggac gaatccacgg gcacagccta
     1261 catggagctg agcagcctga gatctgagga cacggccata tattattgtg cgagagggga
     1321 tacgattttt ggagtgacca tgggatacta cgctatggac gtctggggcc aagggaccac
     1381 ggtcaccgtc tccgcagcct ccaccaaggg cccatcggtc ttccccctgg caccctcctc
     1441 caagagcacc tctgggggca cagcggccct gggctgcctg gtcaaggact acttccccga
     1501 accggtgacg gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc
     1561 tgtcctacag tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag
     1621 cttgggcacc cagacctaca tctgcaacgt gaatcacaag cccagcaaca ccaaggtgga
     1681 caagaaagtt gagcccaaat cttgtgacaa aactagtggc caggccggcc agcaccatca
     1741 ccatcaccat ggcgcatacc cgtacgacgt tccggactac gcttcttagg agggtggtgg
     1801 ctctgagggt ggcggttctg agggtggcgg ctctgaggga ggcggttccg gtggtggctc
     1861 tggttccggt gattttgatt atgaaaagat ggcaaacgct aataaggggg ctatgaccga
     1921 aaatgccgat gaaaacgtgc tacagtctga cgctaaaggc aaacttgatt ctgtcgctac
     1981 tgattacggt gctgctatcg atggtttcat tggtgacgtt tccggccttg ctaatggtaa
     2041 tggtgctact ggtgattttg ctggctctaa ttcccaaatg gctcaagtcg gtgacggtga
     2101 taattcacct ttaatgaata atttccgtca atatttacct tccctccctc aatcggttga
     2161 atgtcgccct tttgtctttg gcgctggtaa accatatgaa ttttctattg attgtgacaa
     2221 aataaactta ttccgtggtg tctttgcgtt tcttttatat gttgccacct ttatgtatgt
     2281 attttctacg tttgctaaca tactgcgtaa taaggagtct taagctagct aattaattta
     2341 agcggccgca gatctgctct ctgaggagga tctgggaaat tgtaagcgtt aatattttgt
     2401 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2461 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2521 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     2581 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     2641 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     2701 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     2761 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     2821 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2881 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2941 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3001 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3061 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3121 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3181 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3241 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3301 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3361 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3421 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3481 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3541 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3601 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3661 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3721 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3781 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     3841 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     3901 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     3961 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4021 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4081 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4141 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4201 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4261 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4321 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4381 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4441 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4501 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4561 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4621 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4681 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4741 agtcagtgag cgaggaagcg gaagag
//
