LOCUS       sequence-349777-        5809 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses IgG affinity protein FB with a TAG stop codon at E25 
            position.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5809)
  AUTHORS   Yu C, Tang J, Loredo A, Chen Y, Jung SY, Jain A, Gordon A, Xiao H
  TITLE     Proximity-Induced Site-Specific Antibody Conjugation.
  JOURNAL   Bioconjug Chem. 2018 Nov 21;29(11):3522-3526. doi: 
            10.1021/acs.bioconjchem.8b00680. Epub 2018 Nov 1.
  PUBMED    30372039
REFERENCE   2  (bases 1 to 5809)
  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"; doi: 
            "10.1021/acs.bioconjchem.8b00680"; journalName: "Bioconjug Chem"; 
            date: "2018-11-21- 21"; volume: "29"; issue: "11"; pages: 
            "3522-3526"
COMMENT     Sequence Label: pET22b-T5-FB-E25TAG
FEATURES             Location/Qualifiers
     source          1..5809
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(79..1161)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1123..1142
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1162..1239)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1445..1464
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1548..1567
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1548..1566
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1567..1591
                     /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        join(1631..1645,1646..1651,1652..1668,1669..1675)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1651..1667
                     /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)."
     protein_bind    1683..1699
                     /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)."
     RBS             1718..1729
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1919..1936
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1962..2056
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2088..2105
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2158..2176)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2172..2219
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2256..2711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2343..2362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2553..2574
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2737..2841
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2842..2910,2911..3702)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3060..3079)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3873..4461
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4362..4381
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4615..4632
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4647..4786
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4712..4731)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4872..4894
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4888..5079)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
       61 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
      121 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
      181 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
      241 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
      301 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
      361 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
      421 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
      481 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
      541 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
      601 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
      661 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
      721 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
      781 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
      841 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
      901 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
      961 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     1021 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     1081 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     1141 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     1201 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     1261 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     1321 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     1381 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     1441 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     1501 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
     1561 tataggggaa ttgtgagcgg ataacaattc ccctctagag tttgacagca ttatcatcga
     1621 tctcgagaaa tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc
     1681 aattgtgagc ggataacaat ttcacacaga attcattaaa gaggagaaat tacatatggt
     1741 ggacaataaa ttcaacaagg agcaacagaa cgccttttat gaaatccttc atttgccaaa
     1801 ccttaactag gaacagagga atgcgtttat tcaatcactt aaggatgatc caagtcaatc
     1861 ggccaacctg ttggcggagg ccaaaaaatt aaatgacgca caagcgccta aaggatccca
     1921 ccaccatcat catcattgag aagcttaatt agctgagctt ggactcctgt tgatagatcc
     1981 agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc gccgggcgtt
     2041 ttttattggt gagaatccaa gctagcttgg cggcggccgc actcgagcac caccaccacc
     2101 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
     2161 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2221 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
     2281 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     2341 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     2401 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     2461 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     2521 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     2581 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     2641 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     2701 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     2761 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     2821 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     2881 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     2941 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     3001 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     3061 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     3121 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     3181 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3241 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3301 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3361 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     3421 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3481 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3541 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3601 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3661 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     3721 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     3781 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     3841 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     3901 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     3961 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4021 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4081 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4141 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4201 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4261 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4321 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     4381 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     4441 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     4501 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     4561 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     4621 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     4681 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4741 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     4801 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4861 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4921 gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg
     4981 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
     5041 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctctg tgtaaggggg
     5101 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
     5161 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
     5221 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
     5281 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
     5341 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
     5401 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
     5461 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
     5521 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
     5581 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     5641 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     5701 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     5761 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgact
//
