LOCUS       sequence-402569-        9564 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Baculovirus transfer vector for expression of huntingtin protein in 
            insect and mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9564)
  TITLE     Unpublished huntingtin plasmids
REFERENCE   2  (bases 1 to 9564)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: huntingtin_V2095-V3138_NbioCflag
FEATURES             Location/Qualifiers
     source          1..9564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      102..556
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(189..208)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     398..419
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        582..686
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(687..755,756..1547)
                     /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(905..924)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1718..2306
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2207..2226
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2460..2477
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2611..2835
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2902..3435)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3624..3629,3630..3646,3647..3652))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4160..4208
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4233..4252
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     enhancer        4313..4692
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4693..4896
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4847..4867
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        4943..5052
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             5087..5131
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     regulatory      7635..7644
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             8312..8335
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             8357..8386
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             8387..8410
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(8500..8520)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(8918..8929)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    9154..9288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(9191..9210)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     9245..9264
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  9317..9482
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
      121 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      181 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      241 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      301 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      361 ttttcgccct ttgacgttgg agtccacgtt cttaatagtg gactcttgtt ccaaactgga
      421 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      481 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      541 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      601 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      661 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      721 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      781 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      841 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      901 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      961 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1021 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     1081 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     1141 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     1201 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     1261 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     1321 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     1381 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     1441 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     1501 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1561 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1621 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1681 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1741 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1801 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1861 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1921 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1981 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     2041 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     2101 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     2161 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     2221 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     2281 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     2341 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     2401 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     2461 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     2521 tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct ggcaaaatcg gttacggttg
     2581 agtaataaat ggatgccctg cgtaagcggg tgtgggcgga caataaagtc ttaaactgaa
     2641 caaaatagat ctaaactatg acaataaagt cttaaactag acagaatagt tgtaaactga
     2701 aatcagtcca gttatgctgt gaaaaagcat actggacttt tgttatggct aaagcaaact
     2761 cttcattttc tgaagtgcaa attgcccgtc gtattaaaga ggggcgtggc caagggcatg
     2821 gtaaagacta tattcgcggc gttgtgacaa tttaccgaac aactccgcgg ccgggaagcc
     2881 gatctcggct tgaacgaatt gttaggtggc ggtacttggg tcgatatcaa agtgcatcac
     2941 ttcttcccgt atgcccaact ttgtatagag agccactgcg ggatcgtcac cgtaatctgc
     3001 ttgcacgtag atcacataag caccaagcgc gttggcctca tgcttcaaga gattgatgag
     3061 cgcggtggca atgccctgcc tccggtgctc gccggagact gcgagatcat agatatagat
     3121 ctcactacgc ggctgctcaa acttgggcag aacgtaagcc gcgagagcgc caacaaccgc
     3181 ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta cggagcaagt tcccgaggta
     3241 atcggagtcc ggctgatgtt gggagtaggt ggctacgtct ccgaactcac gaccgaaaag
     3301 atcaagagca gcccgcatgg atttgacttg gtcagggccg agcctacatg tgcgaatgat
     3361 gcccatactt gagccaccta actttgtttt agggcgactg ccctgctgcg taacatcgtt
     3421 gctgctgcgt aacatcgttg ctgctccata acatcaaaca tcgacccacg gcgtaacgcg
     3481 cttgctgctt ggatgcccga ggcatagact gtacaaaaaa acagtcataa caagccatga
     3541 aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa ggttctggac cagttgcgtg
     3601 agcgcatacg ctacttgcat tacagtttac gaaccgaaca ggcttatgtc aactgggttc
     3661 gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac cttgggcagc agcgaagtcg
     3721 aggcatttct gtcctggctg gcgaacgagc gcaaggtttc ggtctccacg catcgtcagg
     3781 cattggcggc cttgctgttc ttctacggca aggtgctgtg cacggatctg cccttgcttc
     3841 aggagatcgg tagacctcgg ccgtcgcggc gcttgccggt ggtgctgacc ccggatgaag
     3901 tggttcgcat cctcggtttt ctggaaggcg agcatcgttt gttcgcccag gactctagct
     3961 atagttctag tggttggcta cgtacccgta gtggctatgg cagggcttgc cgccccgacg
     4021 ttggctgcga gccctgggcc ttcacccgaa cttgggggtt ggggtgggga aaaggaagaa
     4081 acgcgggcgt attggtccca atggggtctc ggtggggtat cgacagagtg ccagccctgg
     4141 gaccgaaccc cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt
     4201 tattgccgtc atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct
     4261 cccccatctc ccggtaccgc atgcggatct gtacgggcca gatatacgcg ttgacattga
     4321 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     4381 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     4441 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     4501 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     4561 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     4621 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     4681 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
     4741 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
     4801 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
     4861 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact
     4921 gcttactggc ttatcgaata cggaccttta attcaaccca acacaatata ttatagttaa
     4981 ataagaatta ttatcaaatc atttgtatat taattaaaat actatactgt aaattacatt
     5041 ttatttacaa tcactcgacg gatccttaag aaggagatat actatgggac tcaatgacat
     5101 tttcgaagcc cagaagatag agtggcatga gggtggatct ggcggctcag ggggtagcgg
     5161 gggctccgtg agtccggaca aagactggta cgttcatctt gtcaaatccc agtgttggac
     5221 caggtcagat tctgcactgc tggaaggtgc agagctggtg aatcggattc ctgctgaaga
     5281 tatgaatgcc ttcatgatga actcggagtt caacctaagc ctgctagctc catgcttaag
     5341 cctagggatg agtgaaattt ctggtggcca gaagagtgcc ctttttgaag cagcccgtga
     5401 ggtgactctg gcccgtgtga gcggcaccgt gcagcagctc cctgctgtcc atcatgtctt
     5461 ccagcccgag ctgcctgcag aaccggcggc ctactggagc aagttgaatg atctgtttgg
     5521 ggatgctgca ctgtatcagt ccctgcccac tctggcccgg gccctggcac agtacctggt
     5581 ggtggtctcc aaactgccca gtcatttgca ccttcctcct gagaaagaga aggacattgt
     5641 gaaattcgtg gtggcaaccc ttgaggccct gtcctggcat ttgatccatg agcagatccc
     5701 gctgagtctg gatctccagg cagggctgga ctgctgctgc ctggccctgc agctgcctgg
     5761 cctctggagc gtggtctcct ccacagagtt tgtgacccac gcctgctccc tcatccactg
     5821 tgtgcacttc atcctggagg ccgttgcagt gcagcctgga gagcagcttc ttagtccaga
     5881 aagaaggaca aataccccaa aagccatcag cgaggaggag gaggaagtag atccaaacac
     5941 acagaatcct aagtatatca ctgcagcctg tgagatggtg gcagaaatgg tggagtctct
     6001 gcagtcggtg ttggccttgg gtcataaaag gaatagcggc gtgccggcgt ttctcacgcc
     6061 attgctcagg aacatcatca tcagcctggc ccgcctgccc cttgtcaaca gctacacacg
     6121 tgtgccccca ctggtgtgga agcttggatg gtcacccaaa ccgggagggg attttggcac
     6181 agcattccct gagatccccg tggagttcct ccaggaaaag gaagtcttta aggagttcat
     6241 ctaccgcatc aacacactag gctggaccag tcgtactcag tttgaagaaa cttgggccac
     6301 cctccttggt gtcctggtga cgcagcccct cgtgatggag caggaggaga gcccaccaga
     6361 agaagacaca gagaggaccc agatcaacgt cctggccgtg caggccatca cctcactggt
     6421 gctcagtgca atgactgtgc ctgtggccgg caacccagct gtaagctgct tggagcagca
     6481 gccccggaac aagcctctga aagctctcga caccaggttt gggaggaagc tgagcattat
     6541 cagagggatt gtggagcaag agattcaagc aatggtttca aagagagaga atattgccac
     6601 ccatcattta tatcaggcat gggatcctgt cccttctctg tctccggcta ctacaggtgc
     6661 cctcatcagc cacgagaagc tgctgctgca gatcaacccc gagcgggagc tggggagcat
     6721 gagctacaaa ctcggccagg tgtccataca ctccgtgtgg ctagggaaca gcatcacacc
     6781 cctgagggag gaggaatggg acgaggaaga ggaggaggag gccgacgccc ctgcaccttc
     6841 gtcaccaccc acgtctccag tcaactccag gaaacaccgg gctggagttg acatccactc
     6901 ctgttcgcag tttttgcttg agttgtacag ccgctggatc ctgccgtcca gctcagccag
     6961 gaggaccccg gccatcctga tcagtgaggt ggtcagatcc cttctagtgg tctcagactt
     7021 gttcaccgag cgcaaccagt ttgagctgat gtatgtgacg ctgacagaac tgcgaagggt
     7081 gcacccttca gaagacgaga tcctcgctca gtacctggtg cctgccacct gcaaggcagc
     7141 tgccgtcctt gggatggaca aggccgtggc ggagcctgtc agccgcctgc tggagagcac
     7201 gctcaggagc agccacctgc ccagcagggt tggagccctg cacggcatcc tctatgtgct
     7261 ggagtgcgac ctgctggacg acactgccaa gcagctcatc ccggtcatca gcgactatct
     7321 cctctccaac ctgaaaggga tcgcccactg cgtgaacatt cacagccagc agcacgtact
     7381 ggtcatgtgt gccactgcgt tttacctcat tgagaactat cctctggacg tagggccgga
     7441 attttcagca tcaataatac agatgtgtgg ggtgatgctg tctggaagtg aggagtccac
     7501 cccctccatc atttaccact gtgccctcag aggcctggag cgcctcctgc tctctgagca
     7561 gctctcccgc ctggatgcag aatcgctggt caagctgagt gtggacagag tgaacgtgca
     7621 cagcccgcac cgggccatgg cggctctggg cctgatgctc acctgcatgt acacaggaaa
     7681 ggagaaagtc agtccgggta gaacttcaga ccctaatcct gcagcccccg acagcgagtc
     7741 agtgattgtt gctatggagc gggtatctgt tctttttgat aggatcagga aaggctttcc
     7801 ttgtgaagcc agagtggtgg ccaggatcct gccccagttt ctagacgact tcttcccacc
     7861 ccaggacatc atgaacaaag tcatcggaga gtttctgtcc aaccagcagc cataccccca
     7921 gttcatggcc accgtggtgt ataaggtgtt tcagactctg cacagcaccg ggcagtcgtc
     7981 catggtccgg gactgggtca tgctgtccct ctccaacttc acgcagaggg ccccggtcgc
     8041 catggccacg tggagcctct cctgcttctt tgtcagcgcg tccaccagcc cgtgggtcgc
     8101 ggcgatcctc ccacatgtca tcagcaggat gggcaagctg gagcaggtgg acgtgaacct
     8161 tttctgcctg gtcgccacag acttctacag acaccagata gaggaggagc tcgaccgcag
     8221 ggccttccag tctgtgcttg aggtggttgc agccccagga agcccatatc accggctgct
     8281 gacttgttta cgaaatgtcg gcgggtccgg ggattacaag gatgacgacg ataagtgaaa
     8341 cctctacttc caatcgcacc atcatcacca tcaccatcac caccatgatt acaaggatga
     8401 cgacgataag tgaggcggcc gccaacttga tatcaagctt atcgataatc aacctctgga
     8461 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     8521 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     8581 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     8641 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     8701 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     8761 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     8821 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     8881 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     8941 tccttcccgc gggcgccccc ctcctcacgg cgagcgaagc ttgtcgagaa gtactagagg
     9001 atctctagag cctgcagtct cgacaagctt gtcgagaagt actagaggat cataatcagc
     9061 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     9121 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     9181 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     9241 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctg atcactgctt
     9301 gagcctagga gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt
     9361 aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct
     9421 aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca
     9481 cagcggggca tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga
     9541 caaaccgtca tcttcggcta cttt
//
