LOCUS       sequence-6773-ex        8100 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 8100)
  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 8100)
  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: pDW445
FEATURES             Location/Qualifiers
     source          1..8100
                     /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(2373..2417)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     promoter        complement(2436..2545)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        2670..2761
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2686..2703
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2810..3775
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAEQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACVAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     3'UTR           4263..4635
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(5318..5335)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(5712..5730)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5713..5730)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5811..5830)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5930..5952
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5990..6008)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6076..6180
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6181..6249,6250..7041)
                     /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(6399..6418)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7212..7800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7701..7720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7954..7971
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8058..8077
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8058..8076
                     /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 tctggccaga
     2341 attctagagg gcccgggcgc gccaccagta tcttcatgcc attcaatttt ttgtgcttca
     2401 aagatatcat tcaggcctcc tgccatggtg cgtacgattg taaataaaat gtaatttaca
     2461 gtatagtatt ttaattaata tacaaatgat ttgataataa ttcttattta actataatat
     2521 attgtgttgg gttgaattaa aggtcccggc atcctcaaat gcataatttc atagtccccc
     2581 ttgttgtaag tgatgcgtat ttctgaatct ttgtaaaata gcacacaaga ctccaacgcg
     2641 tttggcgttt tattttcttg ctcgaggata tcatggagat aattaaaatg ataaccatct
     2701 cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa aacctataaa
     2761 tattccggat tattcatacc gtcccaccat cgggcgtgct agcggatcca tgaaggataa
     2821 caccgtgcca ctgaaattga ttgccctgtt agcgaacggt gaatttcact ctggcgagca
     2881 gttgggtgaa acgctgggaa tgagccgggc ggctattaat aaacacattc agacactgcg
     2941 tgactggggc gttgatgtct ttaccgttcc gggtaaagga tacagcctgc ctgagcccat
     3001 ccagttactt aatgctgaac agatattggg tcagctggat ggcggtagtg tagccgtgct
     3061 gccagttatt gactccacga atcagtacct tcttgatcgt atcggagagc ttaaatcggg
     3121 cgatgcctgt gttgcagaat accagcaggc tggccgtggt cgccgggggc ggaaatggtt
     3181 ttcgcctttt ggcgcaaact tatatttgtc gatgttctgg cgtctggaac aaggcccggc
     3241 ggcggcgatt ggtttaagtc tggttatcgg tatcgtgatg gcggaagtat tacgcaagct
     3301 gggagcagat aaagttcgtg tcaaatggcc taatgacctc tatctgcagg atcgcaagct
     3361 ggcaggcatt cttgtggagc tgactggcaa aactggcgat gcggcgcaaa tagtcattgg
     3421 agccgggatc aacatggcaa tgcgccgtgt tgaagagagt gtcgttaatc aggggtggat
     3481 cacgctgcag gaagcgggga tcaatctcga tcgtaatacg ttggcggcca tgctaatacg
     3541 tgaattacgt gctgcgttgg aactcttcga acaagaagga ttggcacctt atctgtcgcg
     3601 ctgggaaaag ctggataatt ttattaatcg cccagtgaaa cttatcattg gtgataaaga
     3661 aatatttggc atttcacgcg gaatagacaa acagggggct ttattacttg agcaggatgg
     3721 aataataaaa ccctggatgg gcggtgaaat atccctgcgt agtgcagaaa aataagcttg
     3781 tcgttggatg gaaaggaaaa gagttctaca gggaaacttg gacccgcttc atggaagaca
     3841 gcttccccat tgttaacgac caagaagtga tggatgtttt ccttgttgtc aacatgcgtc
     3901 ccactagacc caaccgttgt tacaaattcc tggcccaaca cgctctgcgt tgcgaccccg
     3961 actatgtacc tcatgacgtg attaggatcg tcgagccttc atgggtgggc agcaacaacg
     4021 agtaccgcat cagcctggct aagaagggcg gcggctgccc aataatgaac cttcactctg
     4081 agtacaccaa ctcgttcgaa cagttcatcg atcgtgtcat ctgggagaac ttctacaagc
     4141 ccatcgttta catcggtacc gactctgctg aagaggagga aattctcctt gaagtttccc
     4201 tggtgttcaa agtaaaggag tttgcaccag acgcacctct gttcactggt ccggcgtatt
     4261 aaaacacgat acattgttat tagtacattt attaagcgct agattctgtg cgttgttgat
     4321 ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt
     4381 atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat
     4441 cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac
     4501 cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta
     4561 gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt
     4621 cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg tacaattgac
     4681 tcgacgtaaa cacgttaaat aaagctagct tggacatatt taacatcggg cgtgttagct
     4741 ttattaggcc gattatcgtc gtcgtcccaa ccctcgtcgt tagaagttgc ttccgaagac
     4801 gattttgcca tagccacacg acgcctatta attgtgtcgg ctaacacgtc cgcgatcaaa
     4861 tttgtagttg agctttttgg aattatttct gattgcgggc gtttttgggc gggtttcaat
     4921 ctaactgtgc ccgattttaa ttcagacaac acgttagaaa gcgatggtgc aggcggtggt
     4981 aacatttcag acggcaaatc tactaatggc ggcggtggtg gagctgatga taaatctacc
     5041 atcggtggag gcgcaggcgg ggctggcggc ggaggcggag gcggaggtgg tggcggtgat
     5101 gcagacggcg gtttaggctc aaatgtctct ttaggcaaca cagtcggcac ctcaactatt
     5161 gtactggttt cgggcgccgt ttttggtttg accggtctga gacgagtgcg atttttttcg
     5221 tttctaatag cttccaacaa ttgttgtctg tcgtctaaag gtgcagcggg ttgaggttcc
     5281 gtcggcattg gtggagcggg cggcaattca gacatcgatg gtggtggtgg tggtggaggc
     5341 gctggaatgt taggcacggg agaaggtggt ggcggcggtg ccgccggtat aatttgttct
     5401 ggtttagttt gttcgcgcac gattgtgggc accggcgcag gcgccgctgg ctgcacaacg
     5461 gaaggtcgtc tgcttcgagg cagcgcttgg ggtggtggca attcaatatt ataattggaa
     5521 tacaaatcgt aaaaatctgc tataagcatt gtaatttcgc tatcgtttac cgtgccgata
     5581 tttaacaacc gctcaatgta agcaattgta ttgtaaagag attgtctcaa gctccgcacg
     5641 ccgataacaa gccttttcat ttttactaca gcattgtagt ggcgagacac ttcgctgtcg
     5701 tcgactcgag ttctatagtg tcacctaaat cgtatgtgta tgatacataa ggttatgtat
     5761 taattgtagc cgcgttctaa cgacaatatg tccatatggt gcactctcag tacaatctgc
     5821 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     5881 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5941 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gaggaaaggg cctcgtgata
     6001 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     6061 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6121 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     6181 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     6241 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6301 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6361 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6421 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6481 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6541 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6601 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6661 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6721 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6781 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6841 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6901 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6961 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     7021 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     7081 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     7141 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     7201 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     7261 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7321 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     7381 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7441 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7501 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7561 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     7621 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7681 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7741 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7801 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7861 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7921 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7981 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcaggtta
     8041 acctggctta tcgaaattaa tacgactcac tatagggaga ccggcagatc gatctgtcga
//
