LOCUS       pYY166 pcDNA3.4-His-MBP-hAID        7848 bp DNA     circular SYN 10-DEC-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7848)
  AUTHORS   Yang
  TITLE     Direct Submission
  JOURNAL   Exported Dec 10, 2024 from SnapGene 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7848
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        50..429
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        430..633
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    729
                     /label=MCS
                     /note="multiple cloning site"
     primer_bind     750..797
                     /label=MBP-F
     CDS             750..773
                     /codon_start=1
                     /label=8XHis
                     /translation="HHHHHHHH"
     CDS             774..1874
                     /codon_start=1
                     /label=MBP
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1850..1896
                     /label=MBP-Vector-F
     primer_bind     complement(1851..1895)
                     /label=MBP-R
     CDS             1881..1904
                     /codon_start=1
                     /label=3C
                     /translation="LEVLFQGP"
     CDS             1911..2504
                     /codon_start=1
                     /label=hAID
                     /translation="MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLD
                     FGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNL
                     SLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWE
                     GLHENSVRLSRQLRRILLPLYEVDDLRDAFRTLGL"
     misc_feature    1929..1982
                     /label=AID antibody recognition region
     misc_feature    2247..2279
                     /label=specific loop
     misc_feature    2463..2504
                     /label=AID antibody recognition region
     CDS             2517..2537
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV Site
                     /translation="ENLYFQG"
     CDS             2538..2555
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(2570..2618)
                     /label=p3.4-hc_REV
     primer_bind     complement(2577..2592)
                     /label=HC-SEQ_REV
     CDS             2586..2609
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     misc_feature    2624..3212
                     /label=WPRE
                     /note="woodchuck posttranscriptional regulatory element"
     primer_bind     complement(2690..2707)
                     /label=WPRE-r
     CDS             complement(3095..3106)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3278..3326
                     /label=HSV TK poly(A) signal
                     /note="herpesvirus thymidine kinase polyadenylation signal"
     rep_origin      3528..3956
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3970..4299
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4150..4285
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4366..5160
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5336..5457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5506..5522)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5530..5546
                     /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        complement(5554..5584)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5599..5620
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5908..6496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6667..7527)
                     /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"
     promoter        complement(7528..7632)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagg atcgaggcgc gccatgggcc atcatcatca ccatcaccat cacatgaaaa
      781 tcgaagaagg taaactggta atctggatta acggcgataa aggctataac ggtctcgctg
      841 aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata
      901 aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac attatcttct
      961 gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg
     1021 acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca
     1081 agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc
     1141 tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga
     1201 aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg
     1261 ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg
     1321 gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca
     1381 aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa
     1441 cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt
     1501 atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc
     1561 tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa
     1621 actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg
     1681 tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccactatgg
     1741 aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg
     1801 ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga
     1861 aagacgcgca gacttcaggt ctggaagtgc tgtttcaggg cccgggatcc atggacagcc
     1921 tcttgatgaa ccggaggaag tttctttacc aattcaaaaa tgtccgctgg gctaagggtc
     1981 ggcgtgagac ctacctgtgc tacgtagtga agaggcgtga cagtgctaca tccttttcac
     2041 tggactttgg ttatcttcgc aataagaacg gctgccacgt ggaattgctc ttcctccgct
     2101 acatctcgga ctgggaccta gaccctggcc gctgctaccg cgtcacctgg ttcacctcct
     2161 ggagcccctg ctacgactgt gcccgacatg tggccgactt tctgcgaggg aaccccaacc
     2221 tcagtctgag gatcttcacc gcgcgcctct acttctgtga ggaccgcaag gctgagcccg
     2281 aggggctgcg gcggctgcac cgcgccgggg tgcaaatagc catcatgacc ttcaaagatt
     2341 atttttactg ctggaatact tttgtagaaa accatgaaag aactttcaaa gcctgggaag
     2401 ggctgcatga aaattcagtt cgtctctcca gacagcttcg gcgcatcctt ttgcccctgt
     2461 atgaggttga tgacttacga gacgcatttc gtactttggg actttgagcg gccgctgaaa
     2521 acctttactt tcagggatgc tgcccgggtt gttgtgatta caaggacgac gacgatgcag
     2581 ctagacatca ccatcaccat caccatcact aagatatctc gacaatcaac ctctggatta
     2641 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2701 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2761 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2821 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2881 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2941 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3001 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     3061 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3121 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3181 gagtcggatc tccctttggg ccgcctcccc gcctggaaac gggggaggct aactgaaaca
     3241 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     3301 acgcacgggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     3361 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     3421 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     3481 ccctgccata gcagatctgc gcagctgggg ctctaggggg tatccccacg cgccctgtag
     3541 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3601 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3661 tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca
     3721 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3781 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3841 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg
     3901 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3961 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     4021 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     4081 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4141 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4201 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     4261 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     4321 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4381 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4441 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4501 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4561 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4621 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4681 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4741 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4801 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4861 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4921 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4981 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     5041 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     5101 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     5161 gcgggactct ggggttcgcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5221 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5281 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5341 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5401 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5461 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5521 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5581 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5641 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5701 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5761 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5821 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5881 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5941 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6001 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6061 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     6121 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6181 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6241 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6301 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6361 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6421 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6481 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6541 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6601 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6661 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6721 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6781 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6841 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6901 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6961 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7021 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7081 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7141 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7201 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7261 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7321 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7381 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7441 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7501 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7561 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7621 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct
     7681 cccgatcccc tatggtcgac tctcagtaca atctgctctg atgccgcata gttaagccag
     7741 tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct
     7801 acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagg
//
