LOCUS       sequence-151430-        6160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  constitutively expresses FLAG-HA-tagged RNAseH1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6160)
  TITLE     RNAseH1 plasmids (unpublished)
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6160)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pFRT-DestFLAGHA_RNAseH1
FEATURES             Location/Qualifiers
     source          1..6160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    270..391
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(281..300)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     335..354
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(521..1543)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(521..1543)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(1551..1598)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     1730..1749
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    1882..2106
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2095..2112
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2150..2191)
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     protein_bind    2244..2268
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3179..3203)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(3219..3245)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(3246..3269)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3269..3278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3342..3361)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3343..3361)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3406..3609)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3435..3455)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3610..3989
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     4161..4180
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4255..4359
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4360..4428,4429..5220)
                     /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(4578..4597)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5391..5979
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5880..5899
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6133..6150
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctct
      241 agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac
      301 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
      361 tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat
      421 ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt
      481 catagaaggc ggcggtggaa tcgaaatctc gtagtacgtg ctattccttt gccctcggac
      541 gagtgctggg gcgtcggttt ccactatcgg cgagtacttc tacacagcca tcggtccaga
      601 cggccgcgct tctgcgggcg atttgtgtac gcccgacagt cccggctccg gatcggacga
      661 ttgcgtcgca tcgaccctgc gcccaagctg catcatcgaa attgccgtca accaagctct
      721 gatagagttg gtcaagacca atgcggagca tatacgcccg gagccgcggc gatcctgcaa
      781 gctccggatg cctccgctcg aagtagcgcg tctgctgctc catacaagcc aaccacggcc
      841 tccagaagaa gatgttggcg acctcgtatt gggaatcccc gaacatcgcc tcgctccagt
      901 caatgaccgc tgttatgcgg ccattgtccg tcaggacatt gttggagccg aaatccgcgt
      961 gcacgaggtg ccggacttcg gggcagtcct cggcccaaag catcagctca tcgagagcct
     1021 gcgcgacgga cgcactgacg gtgtcgtcca tcacagtttg ccagtgatac acatggggat
     1081 cagcaatcgc gcatatgaaa tcacgccatg tagtgtattg accgattcct tgcggtccga
     1141 atgggccgaa cccgctcgtc tggctaagat cggccgcagc gatcgcatcc atggcctccg
     1201 cgaccggctg cagaacagcg ggcagttcgg tttcaggcag gtcttgcaac gtgacaccct
     1261 gtgcacggcg ggagatgcaa taggtcaggc tctcgctgaa ttccccaatg tcaagcactt
     1321 ccggaatcgg gagcgcggcc gatgcaaagt gccgataaac ataacgatct ttgtagaaac
     1381 catcggcgca gctatttacc cgcaggacat atccacgccc tcctacatcg aagctgaaag
     1441 cacgagattc ttcgccctcc gagagctgca tcaggtcgga gacgctgtcg aacttttcga
     1501 tcagaaactt ctcgacagac gtcgcggtga gttcaggctt tttggccaag gaagttccta
     1561 tactttctag agaataggaa cttcggaata ggaacttcta ggtacgtgaa ccatcaccct
     1621 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     1681 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     1741 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     1801 cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atacccccta gagccccagc
     1861 tggttctttc cgcctcagaa gccatagagc ccaccgcatc cccagcatgc ctgctattgt
     1921 cttcccaatc ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc
     1981 tactcagaca atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac
     2041 cttccagggt caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag
     2101 gcacagtcga ggctgatcag cgggtttaaa cgggccctct agactcgagc gtagaatcga
     2161 gaccgaggag agggttaggg ataggcttac cttcgaaccg cgggccctct agacttgagc
     2221 ggccgccact gtgctggata tcaaccactt tgtacaagaa agctgggtct agatatctcg
     2281 agtgcggccg ctcagtcttc cgattgttta gctccttctc tggctaatct gtcagcttct
     2341 tcattgccta taaatcccga atgaccagga acatgcatcc actgaatgtc catcccctgg
     2401 gtaagcctct ccagtgccac aaagtcctct ttgttgatca cctctttccc tgcacttgtc
     2461 ttccacccat ttttcttcca accttgaacc cagttagtta taccatttat cgtaaacata
     2521 ctgtctgtat acagaaccag tttattgatg ttttgagtct ttgcttgttc aatggctttg
     2581 caggctgcat gaatttccgc tctttggttt gtctgccgcc caggaagtct aatgcctaca
     2641 tttaaaggat ggcccggccc ccagtaaacg ccgattcctg ctcgcggcct tctacgccca
     2701 ttactggagc agcagccatc agtgtagacg acgacgaagt ctcccatgta ggaaaacgtg
     2761 tctctgctaa ctggaggcgc cggctccacg ctcggcttca tgtgctttgc atacggctct
     2821 gcgctttcat gtccatctcc atccagtggc tcacggagtc gcttgctggc tttcgcctcc
     2881 gattcttgtc catgttgatt ttcatgccct tctgaaactt ccgggcttgc agatttcctg
     2941 acaaaggccc aggcctcatc ctctgtggca aacttcttaa atctggcagc aggaaaccgg
     3001 tccacctgtg ctctgcactc attccaggtc agaaagaccc cggtcttgcg gcccctcctc
     3061 acggcataga acatcccgaa cccgcgagag ccgcggcggc agggcaaggc ggccaaggcg
     3121 actctgtggg ccaggaacag aagccagctc atgtcgactg aattggttcc catggtggag
     3181 cctgcttttt tgtacaaact tgttgatatc tggttcaaag cgtaatcggg cacgtcataa
     3241 gggtacttgt catcgtcgtc cttgtagtcc atggcggcgg cgatcagaat tccaccacac
     3301 tatcgatgga tcgatccgag ctcgctagcc agcttgggtc tccctatagt gagtcgtatt
     3361 aatttcgata agccagtaag cagtgggttc tctagttagc cagagagctc tgcttatata
     3421 gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca
     3481 ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg
     3541 gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa
     3601 ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc
     3661 ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat
     3721 agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa
     3781 atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt
     3841 cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt
     3901 aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta
     3961 ctattaataa ctagtcaata atcaatgtca acgcgtatat ctggcccgta catcgcgaag
     4021 cagcgcaaaa cgcctaaccc taagcagatt cttcatgcaa ttgtcggtca agccttgcct
     4081 tgttgtagct taaattttgc tcgcgcacta ctcagcgacc tccaacacac aagcagggag
     4141 cagatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagggg
     4201 atcgggagat ctcccgatcc gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     4261 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4321 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4381 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4441 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4501 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4561 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4621 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4681 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4741 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4801 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4861 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4921 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4981 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5041 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5101 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5161 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5221 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5281 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5341 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5401 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5461 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5521 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5581 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5641 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5701 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5761 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5821 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5881 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5941 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6001 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6061 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6121 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
