LOCUS       sequence-469462-        7968 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculovirus expression for structure determination. May not contain 
            entire coding region of gene..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7968)
  TITLE     Arrowsmith Lab SGC Plasmids
REFERENCE   2  (bases 1 to 7968)
  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"
COMMENT     Sequence Label: NL63_NSP13_BV_BIO
FEATURES             Location/Qualifiers
     source          1..7968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..455
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(88..107)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     297..318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        481..585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(586..654,655..1446)
                     /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(804..823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1617..2205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2106..2125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2359..2376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2510..2734
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2801..3334)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3504..3509,3510..3526,3527..3532))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4040..4088
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4113..4132
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4159..5124)
                     /codon_start=1
                     /gene="birA"
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             complement(4162..5124)
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRK
                     LGADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQG
                     WITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIG
                     DKEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     promoter        complement(5161..5270)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5294..5385
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5310..5327
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5436..5480
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     RBS             5963..5971
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7305..7322
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    7457..7591
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7494..7513)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7548..7567
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7620..7785
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
       61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
      241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc ttaatagtgg
      301 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
      361 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
      421 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
      481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
      541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     1021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     1081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     1141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     1201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     1261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     1321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     1381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     1441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1741 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     2041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2281 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2341 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg
     2401 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagaccagcc gcgtaacctg
     2461 gcaaaatcgg ttacggttga gtaataaatg gatgccctgc gtaagcgggt gtgggcggac
     2521 aataaagtct taaactgaac aaaatagatc taaactatga caataaagtc ttaaactaga
     2581 cagaatagtt gtaaactgaa atcagtccag ttatgctgtg aaaaagcata ctggactttt
     2641 gttatggcta aagcaaactc ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag
     2701 gggcgtggcc aagggcatgg taaagactat attcgcggcg ttgtgacaat ttaccgaaca
     2761 actccgcggc cgggaagccg atctcggctt gaacgaattg ttaggtggcg gtacttgggt
     2821 cgatatcaaa gtgcatcact tcttcccgta tgcccaactt tgtatagaga gccactgcgg
     2881 gatcgtcacc gtaatctgct tgcacgtaga tcacataagc accaagcgcg ttggcctcat
     2941 gcttgaggag attgatgagc gcggtggcaa tgccctgcct ccggtgctcg ccggagactg
     3001 cgagatcata gatatagatc tcactacgcg gctgctcaaa cttgggcaga acgtaagccg
     3061 cgagagcgcc aacaaccgct tcttggtcga aggcagcaag cgcgatgaat gtcttactac
     3121 ggagcaagtt cccgaggtaa tcggagtccg gctgatgttg ggagtaggtg gctacgtctc
     3181 cgaactcacg accgaaaaga tcaagagcag cccgcatgga tttgacttgg tcagggccga
     3241 gcctacatgt gcgaatgatg cccatacttg agccacctaa ctttgtttta gggcgactgc
     3301 cctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3361 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3421 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3481 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3541 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3601 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3661 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3721 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3781 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3841 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     3901 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     3961 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4021 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4081 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4141 cccccatctc ccggtacttc atttttctgc actacgcagg gatatttcac cgcccatcca
     4201 gggttttatt attccatcct gctcaagtaa taaagccccc tgtttgtcta ttccgcgtga
     4261 aatgccaaat atttctttat caccaatgat aagtttcact gggcgattaa taaaattatc
     4321 cagcttttcc cagcgcgaca gataaggtgc caatccttct tgttcgaaga gttccaacgc
     4381 agcacgtaat tcacgtatta gcatggccgc caacgtatta cgatcgagat tgatccccgc
     4441 ttcctgcagc gtgatccacc cctgattaac gacactctct tcaacacggc gcattgccat
     4501 gttgatcccg gctccaatga ctatttgcgc cgcatcgcca gttttgccag tcagctccac
     4561 cagaatgcct gccagcttgc gatcctgcag atagaggtca ttaggccatt taacacgaac
     4621 tttatctgca cccagcttgc gtaatacttc cgccatcacg ataccgataa ccagacttaa
     4681 accaatcgcc gccgccgggc cttgttccag acgccagaac atcgacaaat ataagtttgc
     4741 gccaaaaggc gaaaaccatt tccgaccccg gcgaccacgg ccagcctgct ggtattctgc
     4801 aatgcaagca tcgcccgatt taagctctcc gatacgatca agaaggtact gattcgtgga
     4861 gtcaatcact ggcagcacgg ctacactacc gccatccagc tgacccaata tctgtttagc
     4921 attaagtaac tggataggct caggcaggct gtatccttta cccggaacgg taaagacatc
     4981 aacgccccag tcacgcagtg tctgaatgtg tttattaata gccgcccggc tcattcccag
     5041 cgtttcaccc aactgctcgc cagagtgaaa ttcaccgttc gctaacaggg caatcaattt
     5101 cagtggcacg gtgttatcct tcatggctcg agatcccggg tgatcaagtc ttcgtcgagt
     5161 gattgtaaat aaaatgtaat ttacagtata gtattttaat taatatacaa atgatttgat
     5221 aataattctt atttaactat aatatattgt gttgggttga attaaaggtc cgtatactcc
     5281 ggaatattaa tagatcatgg agataattaa aatgataacc atctcgcaaa taaataagta
     5341 ttttactgtt ttcgtaacag ttttgtaata aaaaaaccta taaatattcc ggattattca
     5401 taccgtccca ccatcgggcg cggatcccga tgagcggcct gaacgatatt tttgaagcgc
     5461 agaaaattga atggcatgaa ggcagcgctg gaggttcagg tgcggcgggt ctttgcgtcg
     5521 tatgcggatc tcagacggtt ttaaggtgtg gggattgttt gagaaagccg atgctctgca
     5581 ccaaatgtgc atatgaccac gtttttggaa ccgatcacaa gttcatttta gcgattacgc
     5641 cctacgtttg taatgcgtct ggatgtgggg ttagtgatgt caaaaagctc tatttaggag
     5701 ggcttaatta ctattgtaca aatcataagc cccaactttc tttccctctt tgttcggccg
     5761 gtaacatatt tgggttatac aagaattcag caacagggtc gctcgatgta gaggtgttca
     5821 atcgacttgc gacgtctgac tggacggatg tccgcgatta taaacttgcc aatgacgtta
     5881 aagacacgct cagattattc gctgcggaga ccattaaagc caaggaggag tcggttaaga
     5941 gttcatatgc ttttgccacg cttaaggagg ttgtcggtcc caaagaattg ttgctaagct
     6001 gggaatccgg taaagtgaaa ccgcctctaa accgtaattc ggtcttcact tgctttcaga
     6061 taagtaagga cagcaaattc caaatcggag agtttatttt tgaaaaagtc gaatatggct
     6121 ctgacactgt tacttacaag tccaccgtga ccactaagct agttccaggc atgatatttg
     6181 tactgacaag tcataatgtt cagccgctac gggctccgac aatcgcgaac caggaaaaat
     6241 actcgtccat ttataagtta catccagcgt tcaatgtgag cgatgcatac gctaacctgg
     6301 tcccttacta tcagttgata ggcaaacaga aaatcaccac gatccaaggc ccccctggaa
     6361 gtggtaaatc acattgctct atcggcctag ggctgtacta tccaggtgcg cgaattgtat
     6421 ttgtagcctg cgctcatgct gcggtcgatt cgctgtgtgc aaaggccatg actgtttatt
     6481 caatcgacaa atgcacacgg atcattccag cacgcgccag agtggagtgt tactcagggt
     6541 tcaaacctaa caatactagt gcacagtaca tctttagcac cgtcaatgca ctacccgagt
     6601 gtaacgctga catagtggtt gtcgatgagg tgtccatgtg cacaaactac gatctgtccg
     6661 taatcaacca gcggttatcg tacaaacaca ttgtgtatgt aggcgacccg caacaattgc
     6721 cagccccacg tgtcatgatt acgaaaggtg taatggaacc ggtggattat aacgtcgtca
     6781 cacagcgaat gtgcgccatc ggtccggacg tattcttgca caagtgttat aggtgccctg
     6841 ctgaaatagt taatacggtc tccgagttgg tgtatgaaaa caaattcgtg cccgtgaagc
     6901 cagcaagtaa acaatgcttc aaggtatttt tcaagggcaa cgtgcaggta gacaacggat
     6961 cttccattaa tcggaagcaa ctcgaaatag taaagctttt tctcgtcaag aacccctcct
     7021 ggtcaaaggc agtattcatt tcgccttaca actcgcaaaa ctatgtggcc tcacgttttc
     7081 taggcctcca gattcaaact gtggattcta gtcaggggtc agagtatgac tacgttatct
     7141 acgctcaaac ttcagacacc gcacacgcat gcaatgtaaa ccgctttaat gtagctataa
     7201 caagggccaa aaagggaata ttctgcgtaa tgtgtgacaa gacacttttt gatagcctga
     7261 aatttttcga aataaaacat gctgatctgc acggcggaag tggacatcac caccatcatc
     7321 actgatgaag cttgtcgaga agtactagag gatcataatc agccatacca catttgtaga
     7381 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     7441 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     7501 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     7561 actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta ggagatccga
     7621 accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc gtattagctt
     7681 acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat ccttaaaaac
     7741 tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg gcatttttct
     7801 tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg tcatcttcgg
     7861 ctactttttc tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta ttaatgtttg
     7921 taattgactg aatatcaacg cttatttgca gcctgaatgg cgaatggg
//
