LOCUS pComb3 4030 bp ds-DNA circular SYN 14-JAN-2022
DEFINITION For phage display of antibody libraries by fusion to modified
geneIII..
ACCESSION .
VERSION .
KEYWORDS pComb3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4030)
AUTHORS Barbas CF 3rd, Kang AS, Lerner RA, Benkovic SJ
TITLE Assembly of combinatorial antibody libraries on phage surfaces: the
gene III site.
JOURNAL Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7978-82.
PUBMED 1896445
REFERENCE 2 (bases 1 to 4030)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 14, 2022 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4030
/organism="synthetic DNA construct"
/mol_type="other DNA"
sig_peptide 46..111
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/label=pelB signal sequence
primer_bind 125..141
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 126..142
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 924..945
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 960..990
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 998..1014
/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 1059..1124
/gene="pelB (fragment)"
/product="leader peptide for secretion"
/label=pelB signal sequence
primer_bind complement(1185..1204)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(1186..1204)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(1211..1228)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(1211..1227)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1220..1242)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin complement(1369..1824)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1506..1527)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 1718..1737
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 1851..1955
/gene="bla"
/label=AmpR promoter
CDS 1956..2816
/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(2174..2193)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2987..3575
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 3476..3495
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3729..3746
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 3863..3884
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 3899..3929
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3937..3953
/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)."
primer_bind 3942..3964
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 3961..3977
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 3961..3977
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
promoter 3998..4016
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 caccgcggtg gcggccgcaa attctatttc aaggagacag tcataatgaa atacctattg
61 cctacggcag ccgctggatt gttattactc gctgcccaac cagccatggc ccaggtgaaa
121 ctgctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc cgggggatcc
181 actagtggtg gcggtggctc tccattcgtt tgtgaatatc aaggccaatc gtctgacctg
241 cctcaacctc ctgtcaatgc tggcggcggc tctggtggtg gttctggtgg cggctctgag
301 ggtggtggct ctgagggtgg cggttctgag ggtggcggct ctgagggagg cggttccggt
361 ggtggctctg gttccggtga ttttgattat gaaaagatgg caaacgctaa taagggggct
421 atgaccgaaa atgccgatga aaacgcgcta cagtctgacg ctaaaggcaa acttgattct
481 gtcgctactg attacggtgc tgctatcgat ggtttcattg gtgacgtttc cggccttgct
541 aatggtaatg gtgctactgg tgattttgct ggctctaatt cccaaatggc tcaagtcggt
601 gacggtgata attcaccttt aatgaataat ttccgtcaat atttaccttc cctccctcaa
661 tcggttgaat gtcgcccttt tgtctttggc gctggtaaac catatgaatt ttctattgat
721 tgtgacaaaa taaacttatt ccgtggtgtc tttgcgtttc ttttatatgt tgccaccttt
781 atgtatgtat tttctacgtt tgctaacata ctgcgtaata aggagtctta atcatgccag
841 ttcttttggg tattccgtta ttatgctagc tagtaacacg acaggtttcc cgactggaaa
901 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
961 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
1021 gaattctaaa ctagctagtc gccaaggaga cagtcataat gaaataccta ttgcctacgg
1081 cagccgctgg attgttatta ctcgctgccc aaccagccat ggccgagctc gtcagttcta
1141 gagttaagcg gccgcaatcg agggggggcc cggtacccaa ttcgccctat agtgagtcgt
1201 attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
1261 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
1321 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcaa attgtaagcg
1381 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
1441 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
1501 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
1561 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
1621 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
1681 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
1741 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
1801 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
1861 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
1921 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
1981 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
2041 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
2101 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
2161 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
2221 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
2281 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
2341 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
2401 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
2461 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
2521 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
2581 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
2641 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
2701 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
2761 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
2821 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
2881 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
2941 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
3001 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
3061 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
3121 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
3181 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
3241 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
3301 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
3361 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
3421 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
3481 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
3541 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
3601 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
3661 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
3721 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
3781 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
3841 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
3901 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
3961 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
4021 aaaagctggc
//
