LOCUS       sequence-366448-        6544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express scFv_QLuc in bacteria cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6544)
  AUTHORS   Xiong Y, Zhang Y, Li Z, Reza MS, Li X, Tian X, Ai HW
  TITLE     Engineered Amber-Emitting Nano Luciferase and Its Use for 
            Immunobioluminescence Imaging In Vivo.
  JOURNAL   J Am Chem Soc. 2022 Aug 1. doi: 10.1021/jacs.2c02320.
  PUBMED    35913786
REFERENCE   2  (bases 1 to 6544)
  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"; journalName: "J Am Chem 
            Soc. 2022 Aug 1. doi: 10.1021/jacs.2c02320."
COMMENT     Sequence Label: pET_scFv_QLuc
FEATURES             Location/Qualifiers
     source          1..6544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(365..1447)
                     /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     1409..1428
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1448..1525)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1731..1750
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1834..1853
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1834..1852
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1853..1877
                     /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             1892..1914
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3242..3259
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3312..3330)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3326..3373
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3410..3865
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3497..3516)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3707..3728
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3957..4772)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(3957..4772)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4680..4699
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4764..4783
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4894..5482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5383..5402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5636..5653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5668..5807
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5733..5752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5893..5915
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5909..6100)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
       61 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
      121 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
      181 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
      241 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
      301 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
      361 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
      421 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
      481 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
      541 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
      601 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
      661 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
      721 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
      781 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
      841 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
      901 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
      961 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     1021 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     1081 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     1141 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     1201 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     1261 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     1321 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     1381 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     1441 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     1501 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     1561 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     1621 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     1681 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     1741 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     1801 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     1861 gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc
     1921 atggatgggc agcgatattc agatgaccca gagcccgagc agcctgagcg cgagcgtggg
     1981 cgatcgcgtg accattacct gccgcgcgag ccaggatgtg aacaccgcgg tggcgtggta
     2041 tcagcagaaa ccgggcaaag cgccgaaact gctgatttat agcgcgagct ttctgtatag
     2101 cggcgtgccg agccgcttta gcggcagccg cagcggcacc gattttaccc tgaccattag
     2161 cagcctgcag ccggaagatt ttgcgaccta ttattgccag cagcattata ccaccccgcc
     2221 gacctttggc cagggcacca aagtggaaat taaacgcacc ggcggtggcg gtagcggtgg
     2281 cggtggcagc ggtggtggcg gcagcggcgg cggtggtagc gaagtgcagc tggtggaaag
     2341 cggcggcggc ctggtgcagc cgggcggcag cctgcgcctg agctgcgcgg cgagcggctt
     2401 taacattaaa gatacctata ttcattgggt gcgccaggcg ccgggcaaag gcctggaatg
     2461 ggtggcgcgc atttatccga ccaacggcta tacccgctat gcggatagcg tgaaaggccg
     2521 ctttaccatt agcgcggata ccagcaaaaa caccgcgtat ctgcagatga acagcctgcg
     2581 cgcggaagat accgcggtgt attattgcag ccgctggggc ggcgatggct tttatgcgat
     2641 ggattattgg ggccagggca ccctggtgac cgtgagcagc ggcagcacca gcggcagcgg
     2701 caaaagcagc gaaggcaaag gcaaagtctt cactctcggg gattttgttg gggactggcg
     2761 acagacagcc ggctacaacc aggctcaagt ccttgaacag ggaggtttga ccagtttgtt
     2821 tcagaacctc ggggtgtccg taactccaat ccaaaggatt gtcctgagcg gtgaaaatgg
     2881 gctgaagatc gatatccatg tcatcatccc gtatgaaggt ctgagctgcg accaaatggc
     2941 ccagttcgaa aaaattttta aggtgatata ccgtgtggat gatcatcact ttaaggtgat
     3001 cctgcactat ggcacactgg taatcgacgg ggttacgccg aacatgatcg actatttcgg
     3061 acagccgtat gaaggcatcg ccaagttcga cggcaaaaag atcacagtaa cagggaccct
     3121 gtggaacggc aacacaatta tcgacgagcg cctgatcaac cccgacggct ccctgctgtt
     3181 ccgagtaacc attaacggag tgaccggctg gcgtctgcat gaacgtattc tggcgctcga
     3241 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt
     3301 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3361 gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt
     3421 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3481 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3541 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3601 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3661 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3721 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3781 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3841 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3901 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaat taattcttag
     3961 aaaaactcat cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca
     4021 tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg
     4081 atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt
     4141 aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa
     4201 tccggtgaga atggcaaaag tttatgcatt tctttccaga cttgttcaac aggccagcca
     4261 ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc
     4321 tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc
     4381 aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct
     4441 tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca
     4501 ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt
     4561 ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac
     4621 tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta
     4681 tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa tcgcggccta
     4741 gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa
     4801 gcagacagtt ttattgttca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4861 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5041 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5521 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5581 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     5641 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
     5701 cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5761 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     5821 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     5881 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     5941 cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct
     6001 gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg cttctgataa
     6061 agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc gtgtaagggg
     6121 gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc acgatacggg
     6181 ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa ctggcggtat
     6241 ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc gttaatacag
     6301 atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg aacataatgg
     6361 tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg aagaccattc
     6421 atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta
     6481 tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg
     6541 acag
//
