LOCUS       sequence-6774-ex        8086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8086)
  AUTHORS   Duffy S, Tsao KL, Waugh DS
  TITLE     Site-specific, enzymatic biotinylation of recombinant proteins in 
            Spodoptera frugiperda cells using biotin acceptor peptides.
  JOURNAL   Anal Biochem. 1998 Sep 10. 262(2):122-8.
  PUBMED    9750126
REFERENCE   2  (bases 1 to 8086)
  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: "Anal 
            Biochem. 1998 Sep 10. 262(2):122-8."
COMMENT     Sequence Label: pDW446
FEATURES             Location/Qualifiers
     source          1..8086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_recomb     1265..2092
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             1265..1389
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(1427..2032)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     polyA_signal    2099..2306
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2295..2312
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2343..2387)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     promoter        complement(2422..2531)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        2656..2747
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2672..2689
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2796..3761
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAEQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACVAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     3'UTR           4249..4621
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(5304..5321)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(5698..5716)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5699..5716)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5797..5816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5916..5938
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5976..5994)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6062..6166
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6167..6235,6236..7027)
                     /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(6385..6404)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7198..7786
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7687..7706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7940..7957
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8044..8063
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8044..8062
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcagttcgtt gacgccttcc tccgtgtggc cgaacacgtc gagcgggtgg tcgatgacca
       61 gcggcgtgcc gcacgcgacg cacaagtatc tgtacaccga atgatcgtcg ggcgaaggca
      121 cgtcggcctc caagtggcaa tattggcaaa ttcgaaaata tatacagttg ggttgtttgc
      181 gcatatctat cgtggcgttg ggcatgtacg tccgaacgtt gatttgcatg caagccgaaa
      241 ttaaatcatt gcgattagtg cgattaaaac gttgtacatc ctcgctttta atcatgccgt
      301 cgattaaatc gcgcaatcga gtcaagtgat caaagtgtgg aataatgttt tctttgtatt
      361 cccgagtcaa gcgcagcgcg tattttaaca aactagccat cttgtaagtt agtttcattt
      421 aatgcaactt tatccaataa tatattatgt atcgcacgtc aagaattaac aatgcgcccg
      481 ttgtcgcatc tcaacacgac tatgatagag atcaaataaa gcgcgaatta aatagcttgc
      541 gacgcaacgt gcacgatctg tgcacgcgtt ccggcacgag ctttgattgt aataagtttt
      601 tacgaagcga tgacatgacc cccgtagtga caacgatcac gcccaaaaga actgccgact
      661 acaaaattac cgagtatgtc ggtgacgtta aaactattaa gccatccaat cgaccgttag
      721 tcgaatcagg accgctggtg cgagaagccg cgaagtatgg cgaatgcatc gtataacgtg
      781 tggagtccgc tcattagagc gtcatgttta gacaagaaag ctacatattt aattgatccc
      841 gatgatttta ttgataaatt gaccctaact ccatacacgg tattctacaa tggcggggtt
      901 ttggtcaaaa tttccggact gcgattgtac atgctgttaa cggctccgcc cactattaat
      961 gaaattaaaa attccaattt taaaaaacgc agcaagagaa acatttgtat gaaagaatgc
     1021 gtagaaggaa agaaaaatgt cgtcgacatg ctgaacaaca agattaatat gcctccgtgt
     1081 ataaaaaaaa tattgaacga tttgaaagaa aacaatgtac cgcgcggcgg tatgtacagg
     1141 aagaggttta tactaaactg ttacattgca aacgtggttt cgtgtgccaa gtgtgaaaac
     1201 cgatgtttaa tcaaggctct gacgcatttc tacaaccacg actccaagtg tgtgggtgaa
     1261 gtcatgcatc ttttaatcaa atcccaagat gtgtataaac caccaaactg ccaaaaaatg
     1321 aaaactgtcg acaagctctg tccgtttgct ggcaactgca agggtctcaa tcctatttgt
     1381 aattattgaa taataaaaca attataaatg ctaaatttgt tttttattaa cgatacaaac
     1441 caaacgcaac aagaacattt gtagtattat ctataattga aaacgcgtag ttataatcgc
     1501 tgaggtaata tttaaaatca ttttcaaatg attcacagtt aatttgcgac aatataattt
     1561 tattttcaca taaactagac gccttgtcgt cttcttcttc gtattccttc tctttttcat
     1621 ttttctcctc ataaaaatta acatagttat tatcgtatcc atatatgtat ctatcgtata
     1681 gagtaaattt tttgttgtca taaatatata tgtctttttt aatggggtgt atagtaccgc
     1741 tgcgcatagt ttttctgtaa tttacaacag tgctattttc tggtagttct tcggagtgtg
     1801 ttgctttaat tattaaattt atataatcaa tgaatttggg atcgtcggtt ttgtacaata
     1861 tgttgccggc atagtacgca gcttcttcta gttcaattac accatttttt agcagcaccg
     1921 gattaacata actttccaaa atgttgtacg aaccgttaaa caaaaacagt tcacctccct
     1981 tttctatact attgtctgcg agcagttgtt tgttgttaaa aataacagcc attgtaatga
     2041 gacgcacaaa ctaatatcac aaactggaaa tgtctatcaa tatatagttg ctgatatctc
     2101 cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc
     2161 ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat tttattagga
     2221 aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg ggcaaacaac
     2281 agatggctgg caactagaag gcacagtcga ggctgatcag cgagctcaga tctttacgta
     2341 tcttcatgcc attcaatttt ttgtgcttca aagatatcat tcaggcctcc cgggcgcgcc
     2401 tctagaattc tggccacgta cgattgtaaa taaaatgtaa tttacagtat agtattttaa
     2461 ttaatataca aatgatttga taataattct tatttaacta taatatattg tgttgggttg
     2521 aattaaaggt cccggcatcc tcaaatgcat aatttcatag tcccccttgt tgtaagtgat
     2581 gcgtatttct gaatctttgt aaaatagcac acaagactcc aacgcgtttg gcgttttatt
     2641 ttcttgctcg aggatatcat ggagataatt aaaatgataa ccatctcgca aataaataag
     2701 tattttactg ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt
     2761 cataccgtcc caccatcggg cgtgctagcg gatccatgaa ggataacacc gtgccactga
     2821 aattgattgc cctgttagcg aacggtgaat ttcactctgg cgagcagttg ggtgaaacgc
     2881 tgggaatgag ccgggcggct attaataaac acattcagac actgcgtgac tggggcgttg
     2941 atgtctttac cgttccgggt aaaggataca gcctgcctga gcccatccag ttacttaatg
     3001 ctgaacagat attgggtcag ctggatggcg gtagtgtagc cgtgctgcca gttattgact
     3061 ccacgaatca gtaccttctt gatcgtatcg gagagcttaa atcgggcgat gcctgtgttg
     3121 cagaatacca gcaggctggc cgtggtcgcc gggggcggaa atggttttcg ccttttggcg
     3181 caaacttata tttgtcgatg ttctggcgtc tggaacaagg cccggcggcg gcgattggtt
     3241 taagtctggt tatcggtatc gtgatggcgg aagtattacg caagctggga gcagataaag
     3301 ttcgtgtcaa atggcctaat gacctctatc tgcaggatcg caagctggca ggcattcttg
     3361 tggagctgac tggcaaaact ggcgatgcgg cgcaaatagt cattggagcc gggatcaaca
     3421 tggcaatgcg ccgtgttgaa gagagtgtcg ttaatcaggg gtggatcacg ctgcaggaag
     3481 cggggatcaa tctcgatcgt aatacgttgg cggccatgct aatacgtgaa ttacgtgctg
     3541 cgttggaact cttcgaacaa gaaggattgg caccttatct gtcgcgctgg gaaaagctgg
     3601 ataattttat taatcgccca gtgaaactta tcattggtga taaagaaata tttggcattt
     3661 cacgcggaat agacaaacag ggggctttat tacttgagca ggatggaata ataaaaccct
     3721 ggatgggcgg tgaaatatcc ctgcgtagtg cagaaaaata agcttgtcgt tggatggaaa
     3781 ggaaaagagt tctacaggga aacttggacc cgcttcatgg aagacagctt ccccattgtt
     3841 aacgaccaag aagtgatgga tgttttcctt gttgtcaaca tgcgtcccac tagacccaac
     3901 cgttgttaca aattcctggc ccaacacgct ctgcgttgcg accccgacta tgtacctcat
     3961 gacgtgatta ggatcgtcga gccttcatgg gtgggcagca acaacgagta ccgcatcagc
     4021 ctggctaaga agggcggcgg ctgcccaata atgaaccttc actctgagta caccaactcg
     4081 ttcgaacagt tcatcgatcg tgtcatctgg gagaacttct acaagcccat cgtttacatc
     4141 ggtaccgact ctgctgaaga ggaggaaatt ctccttgaag tttccctggt gttcaaagta
     4201 aaggagtttg caccagacgc acctctgttc actggtccgg cgtattaaaa cacgatacat
     4261 tgttattagt acatttatta agcgctagat tctgtgcgtt gttgatttac agacaattgt
     4321 tgtacgtatt ttaataattc attaaattta taatctttag ggtggtatgt tagagcgaaa
     4381 atcaaatgat tttcagcgtc tttatatctg aatttaaata ttaaatcctc aatagatttg
     4441 taaaataggt ttcgattagt ttcaaacaag ggttgttttt ccgaaccgat ggctggacta
     4501 tctaatggat tttcgctcaa cgccacaaaa cttgccaaat cttgtagcag caatctagct
     4561 ttgtcgatat tcgtttgtgt tttgttttgt aataaaggtt cgacgtcgtt caaaatatta
     4621 tgcgcttttg tatttctttc atcactgtcg ttagtgtaca attgactcga cgtaaacacg
     4681 ttaaataaag ctagcttgga catatttaac atcgggcgtg ttagctttat taggccgatt
     4741 atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt ttgccatagc
     4801 cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg tagttgagct
     4861 ttttggaatt atttctgatt gcgggcgttt ttgggcgggt ttcaatctaa ctgtgcccga
     4921 ttttaattca gacaacacgt tagaaagcga tggtgcaggc ggtggtaaca tttcagacgg
     4981 caaatctact aatggcggcg gtggtggagc tgatgataaa tctaccatcg gtggaggcgc
     5041 aggcggggct ggcggcggag gcggaggcgg aggtggtggc ggtgatgcag acggcggttt
     5101 aggctcaaat gtctctttag gcaacacagt cggcacctca actattgtac tggtttcggg
     5161 cgccgttttt ggtttgaccg gtctgagacg agtgcgattt ttttcgtttc taatagcttc
     5221 caacaattgt tgtctgtcgt ctaaaggtgc agcgggttga ggttccgtcg gcattggtgg
     5281 agcgggcggc aattcagaca tcgatggtgg tggtggtggt ggaggcgctg gaatgttagg
     5341 cacgggagaa ggtggtggcg gcggtgccgc cggtataatt tgttctggtt tagtttgttc
     5401 gcgcacgatt gtgggcaccg gcgcaggcgc cgctggctgc acaacggaag gtcgtctgct
     5461 tcgaggcagc gcttggggtg gtggcaattc aatattataa ttggaataca aatcgtaaaa
     5521 atctgctata agcattgtaa tttcgctatc gtttaccgtg ccgatattta acaaccgctc
     5581 aatgtaagca attgtattgt aaagagattg tctcaagctc cgcacgccga taacaagcct
     5641 tttcattttt actacagcat tgtagtggcg agacacttcg ctgtcgtcga ctcgagttct
     5701 atagtgtcac ctaaatcgta tgtgtatgat acataaggtt atgtattaat tgtagccgcg
     5761 ttctaacgac aatatgtcca tatggtgcac tctcagtaca atctgctctg atgccgcata
     5821 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     5881 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     5941 ttcaccgtca tcaccgaaac gcgcgagagg aaagggcctc gtgatacgcc tatttttata
     6001 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     6061 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     6121 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     6181 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     6241 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     6301 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     6361 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     6421 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     6481 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     6541 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     6601 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     6661 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     6721 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     6781 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     6841 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     6901 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6961 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     7021 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     7081 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     7141 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     7201 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     7261 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     7321 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     7381 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     7441 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     7501 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     7561 caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc ccgaagggag
     7621 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     7681 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     7741 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     7801 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     7861 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     7921 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
     7981 caaaccgcct ctccccgcgc gttggccgat tcattaatgc aggttaacct ggcttatcga
     8041 aattaatacg actcactata gggagaccgg cagatcgatc tgtcga
//
