LOCUS       sequence-355950-        7019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of PolyA-HhR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7019)
  TITLE     Izaurralde lab plasmids
REFERENCE   2  (bases 1 to 7019)
  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: pAc5.1C-FlucV5His6-PolyA-HhR_AB
FEATURES             Location/Qualifiers
     source          1..7019
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..2510
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2530..4182
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MAEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTD
                     AHIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVA
                     PANGIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQ
                     SMYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSH
                     ARDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDY
                     KIQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQG
                     YGLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIM
                     SGYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESI
                     LLQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGV
                     VFVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(2599..2617)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     CDS             4204..4245
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             4255..4272
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4295..4312)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(4369..4388)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4423..4442
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_RNA        4574..4625
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     rep_origin      complement(5146..5734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5226..5245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5905..6696,6697..6765))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6528..6547
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6766..6870)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6938..6956
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 catgaatggc atcaactctg aatcaaatct ttgcagatgc acctacttct catttccact
       61 gtcacatcat ttttccagat ctcgctgcct gttatgtggc ccacaaacca agacacgttt
      121 tatggccatt aaagctggct gatcgtcgcc aaacaccaaa tacataatga atatgtacac
      181 attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga gtaggagaat gcggaggaga
      241 aggagaacga gctgatctag tatctctcca caatccaatg ccaactgacc aactggccat
      301 attcggagca atttgaagcc aatttccatc gcctggcgat cgctccattc ttggctatat
      361 gtttttcacc gttacccggg gccattttca aagactcgtc ggcaagataa gattgtgtca
      421 ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag
      481 tattttgaag aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac
      541 aaaaacatgt tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt
      601 atgtatatac ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac
      661 tgatgatatt ttaagatgat gccagaccaa aaggcttgaa tttctgcgtc ttttgccgaa
      721 cgcagtgcat gtgcaattgt tgttttttgg aatattcaat tttcggactg tccgctttga
      781 tttcagtttc ttggcttatt caaaaagcaa agtaaagcca aaaaagcgag atggcaatac
      841 caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg gcagcggaag gaagggtggg
      901 gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc gacgttggag ccactccttt
      961 gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact cgccggttgt ttttttcttt
     1021 ttatgctgcg ctctctctag cgccatctcg cttacgcatg ctcaacgcac cgcatgttgc
     1081 cgtttccttt tatgcgtcat tttggctcga aataggcaat tatttaaaca aagattagtc
     1141 aacgaaaacg ctaaaataaa taagtctaca atatggttac ttattgccat gtgtgtgcag
     1201 ccaacgatag caacaaaagc aacaacacag gtggctttcc ctctttcact ttttgtttgc
     1261 aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa ttacgctgac aaagagcaga
     1321 cgaagttttg gcgaaaaaca tcaaggcgcc tgatacgaat gcatttgcaa taacaattgc
     1381 gatatttaat attgtttatg aagctgtttg acttcaaaac acacaaaaaa aaaaataaaa
     1441 caaattattt gaaagagaat taggaatcgg acgcttatcg ttagggtaac aacaagaaat
     1501 gcttactgag tcacagcctc tggaaaactg ccgcaagcca gagagagaga gaaaaagagg
     1561 gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc tctctcttcg tctctgttgc
     1621 gcaaacgcat agactgcact gaaaaaatcg attacctatt ttttatgaat gaatatttgc
     1681 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1741 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1801 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1861 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgaatgcc ctactagaag
     1921 atgtgtgttg cacaaaatgt ccctggaata accaatttga agtgcagata gcagtaaacg
     1981 taagctaata tgaatattat ttaactgtaa tgttttaata tcgctggaca ttactaataa
     2041 acccactata aacacatgta catatgtatg ttttggcata caatgagtag ttggggaaaa
     2101 aatgtgtaaa agcaccgtga ccatcacagc ataaagataa ccagctgaag tatcgaatat
     2161 gagtaacccc caaattgaat cacatgccgc aactgatagg acccatggaa gtacactctt
     2221 catggcgata tacaagacac acacaagcac gaacacccag ttgcggagga aattctccgt
     2281 aaatgaaaac ccaatcggcg aacaattcat acccatatat ggtaaaagtt ttgaacgcga
     2341 cttgagagcg gagagcattg cggctgataa ggttttagcg ctaagcgggc tttataaaac
     2401 gggctgcggg accagttttc atatcactac cgtttgagtt cttgtgctgt gtggatactc
     2461 ctcccgacac aaagccgctc catcagccag cagtcgtcta atccagagac cccggatcgg
     2521 ggtaccaaca tggccgaaga cgccaaaaac ataaagaaag gcccggcacc attctatcca
     2581 ctagaggatg gaaccgctgg agagcaactg cataaggcta tgaagagata cgccctggtt
     2641 cctggaacaa ttgcttttac agatgcacat atcgaggtga acatcacgta cgcggaatac
     2701 ttcgaaatgt ccgttcggtt ggcagaagct atgaaacgat atgggctgaa tacaaatcac
     2761 agaatcgtcg tatgcagtga aaactctctt caattcttta tgccggtgtt gggcgcgtta
     2821 tttatcggag ttgcagttgc gcccgcgaac ggcatttata atgaacgtga attgctcaac
     2881 agtatgaaca tttcgcagcc taccgtagtg tttgtttcca aaaaggggtt gcaaaaaatt
     2941 ttgaacgtgc aaaaaaaatt accaataatc cagaaaatta ttatcatgga ttctaaaacg
     3001 gattaccagg gatttcagtc gatgtacacg ttcgtcacat ctcatctacc tcccggtttt
     3061 aatgaatacg attttgtacc agagtccttt gatcgtgaca aaacaattgc actgataatg
     3121 aattcctctg gatctactgg gttacctaag ggtgtggccc ttccgcatag aactgcctgc
     3181 gtcagattct cgcatgccag agatcctatt tttggcaatc aaatcattcc ggatactgcg
     3241 attttaagtg ttgttccatt ccatcacggt tttggaatgt ttactacact cggatatttg
     3301 atatgtggat ttcgagtcgt cttaatgtat agatttgaag aagagctgtt tttacgatcc
     3361 cttcaggatt acaaaattca aagtgcgttg ctagtaccaa ccctattttc attcttcgcc
     3421 aaaagcactc tgattgacaa atacgattta tctaatttac acgaaattgc ttctgggggc
     3481 gcacctcttt cgaaagaagt cggggaagcg gttgcaaaac gcttccatct tccagggata
     3541 cgacaaggat atgggctcac tgagactaca tcagctattc tgattacacc cgagggggat
     3601 gataaaccgg gcgcggtcgg taaagttgtt ccattttttg aagcgaaggt tgtggatctg
     3661 gataccggga aaacgctggg cgttaatcag agaggcgaat tatgtgtcag aggacctatg
     3721 attatgtccg gttatgtaaa caatccggaa gcgaccaacg ccttgattga caaggatgga
     3781 tggctacatt ctggagacat agcttactgg gacgaagacg aacacttctt catagttgac
     3841 cgcttgaagt ctttaattaa atacaaaggg tatcaggtgg cccccgctga attggaatcg
     3901 atattgttac aacaccccaa catcttcgac gcgggcgtgg caggtcttcc cgacgatgac
     3961 gccggtgaac ttcccgccgc cgttgttgtt ttggagcacg gaaagacgat gacggaaaaa
     4021 gagatcgtgg attacgtcgc cagtcaagta acaaccgcga aaaagttgcg cggaggagtt
     4081 gtgtttgtgg acgaagtacc gaaaggtctt accggaaaac tcgacgcaag aaaaatcaga
     4141 gagatcctca taaaggccaa gaagggcgga aagtccaaat tggctcgagg tcacccattc
     4201 gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac cggtcatcat
     4261 caccatcacc attgagttta aacccgctga tcagcctcga ctgtgccttc taaggcctga
     4321 gctcgctgat cagcctcgat cgaggatcca gacatgataa gatacattga tgagtttgga
     4381 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     4441 gctgcggccg catacaaagg atctggctgc taaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4501 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     4561 aaaaaaaaac ccccctgtca ccggatgtgt tttccggtct gatgagtccg tgaggacgaa
     4621 acagggaatt cgatatcaag cttatcgata ccgtcgaccg atgcccttga gagccttcaa
     4681 cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac ttatgactgt
     4741 cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca ttttcggcga
     4801 ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat tcggaatctt
     4861 gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca
     4921 ggccattatc gccggcatgg cggccgacgc gctgggctac gtcttgctgg cgttcgcgac
     4981 gcgaggctgg atggccttcc ccattatgat tcttctcgct tccggcggca tcgggatgcc
     5041 cgcgttgcag gccatgctgt ccaggcaggt agatgacgac catcagggac agcttcaagg
     5101 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5161 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5221 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5281 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5341 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5401 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5461 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5521 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5581 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5641 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5701 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5761 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5821 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5881 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5941 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6001 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6061 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6121 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6181 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg
     6241 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6301 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6361 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6421 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6481 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     6541 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6601 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6661 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6721 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6781 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6841 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6901 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6961 tcgtcttcaa gaattaattc tatattctaa aaacacaaat gatacttcta aaaaaaaat
//
