LOCUS       sequence-337963-        8721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes fly Drosha protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8721)
  TITLE     Zeng Lab Plasmids
REFERENCE   2  (bases 1 to 8721)
  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: p3xFLAG-CMV-fly Drosha
FEATURES             Location/Qualifiers
     source          1..8721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..61
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     53..70
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     54..70
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        231..610
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        611..814
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     765..785
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             844..909
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    4975..5597
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5174..5193)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5621..5641)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5626..5955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5806..5941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5993..6012)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5994..6012)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6026..6042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6026..6042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6039..6061)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6050..6066
                     /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)."
     promoter        complement(join(6074..6080,6081..6098,6099..6104))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6119..6140
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6257..6274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6428..7016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6508..6527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7187..7978,7979..8047))
                     /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     7810..7829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8048..8152)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8179..8634
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8266..8285)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8476..8497
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
       61 gacggccagt gccaagctga tctatacatt gaatcaatat tggcaattag ccatattagt
      121 cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata
      181 tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt
      241 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
      301 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
      361 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
      421 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
      481 tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
      541 ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc
      601 tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc
      661 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
      721 tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag
      781 gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc agaattaacc
      841 atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
      901 gatgacaagc ttatgtacca gccgcctttg ccaccgcctc cagtccagcc tgctcctccg
      961 cccccgccac ctccgccaga agaagatcta tctccgccgg gagtaggtgt tccctctcac
     1021 aactattctt caaatgaatc ccattctcag agctcgaagt ctttggatta cgtctacccg
     1081 gagacaccgg ctccttatgc cagctcagta cctagctatg atccgtacca gcaaccacca
     1141 gcgtacggct acgagggata tgcttacaac gaacaagcac agaagtatgg tggccaggag
     1201 tcccattacc aataccagta cccggcatcc ggatcatcat tcctctatga aagctataaa
     1261 tacccagacc gctatcctgc ttattcgtcc aactaccgac ctccatccga aaggcagagg
     1321 tacacttcca acagctccag tcaaggctac caccattatc ccggctatag ctcgggtagg
     1381 cggtatgaac aaagacatga tcaggaacac cgtcaaattc aggacagtcg atatgcccac
     1441 gagccacgac acggacacta tgcccacagg caggcaaagg ggtcgcaaca cggatactat
     1501 ggatctgcag ccagaaacca agtcagcgat gactattcgc cacgtggcca tcacgaaagg
     1561 gagagaaatg agaccttaga gaagactagg gccaaaccga aggtggagac cgaaagggac
     1621 cgtctgttga gacagtggtg ttctaacttc tgcgaaaaac cagaggacta tgtgaagaag
     1681 atgaacgccc tcagcgaggc ggatgcgcca gtcgaatcct gggtgcgatc atcgccagcg
     1741 gagctctact acgagcgcac caagagcgaa aacgaagtca gaggtcgggc gcgtctgcaa
     1801 aagctgtgca ctctattcga tgaggagctc ctgcagagag caaagcgtgt acgcgagaag
     1861 ctacccgttt acgtgccgcc tccaagaaag gctcgccgcc gtgtttgcaa gcataaacac
     1921 aagtcggaag cctgctcctc gtcctcctca tccgatgacg actccgacga agacgctttt
     1981 aagatcgagc aagactgctg catggaggag ctttcgcgga aggtgcaaca tccgcagcgt
     2041 gtgcacgcgg atctctggca caacgacgcc ggtgagatga acgacggacc tctttgccgc
     2101 tgctcagcca agtcccggcg cattggcatt cggcacggaa tatatccggg cgagactggc
     2161 tacaagttgt gcgatccaaa cagcaacaat gcaggcaagc tgttccacta caggatcagc
     2221 atctcaccgc ccactaactt cctgacaaag acacccacca ttatcaagca tgatgagcac
     2281 gagtttctat tcgagggctt ctcacttctc tcgcatgtgc gtctctccga tctgcccgtc
     2341 tgcaaggtga tccgcttcaa catcgagtac accattgagt acgaagagga gaagatgcca
     2401 gagaacttca ccatccacga gctagacatt tttttcaaat acctgtttca tgaactgcta
     2461 gagctggtag actttaatct aatgcccaac ttaccgtccg gaaacgtcga ggaatcctgt
     2521 ccagcttttc acttctttcc gcgtttcgtc cgcgacctgc cagataatgg aaaggaggtt
     2581 ttggccatgg tcgaggtact ccgctacttg ctggataatt ctgcacagct tgtggaacgg
     2641 cagcaactac tgcatcttaa ccagattagt cagagcgagt ggcaaaacta cgtggacttc
     2701 ataaagggaa tgctggtcac caagccgggt tataagccgt gttcgctacg cgttgaccaa
     2761 ttggacagga ataactccga tttgcccgag tgcgtagatc gcgagactgg aatctcacat
     2821 ccagcaatcg tgcactttgg catttgtcat cctcagctaa gctacgctgg aaatccagag
     2881 taccagaagg cgtggcgaga gtacgttaag taccgtcatc tgatggccaa catgtcgaag
     2941 ccctctttca aggataagcg caagctagag gagaaggagc aacgtcttca ggagatgcga
     3001 actcaggggc gcatgaaacg aaatatcaca gtggcgatca gctcggaggg cttctatcgc
     3061 accggcatta tgtgcgacgt tgtgcagcat gccatgttga ttcctgtcct aactggtcac
     3121 cttcgctttc acaagtcgct ggacctgcta gaggagagta tcgggtaccg ctttaaaaat
     3181 cggtaccttc tccaattggc gctgacgcat ccctcataca aggagaacta cggtaccaat
     3241 ccggatcacg cccgtaattc gctgactaac tgcggaattc gtcagccgga gtacggagat
     3301 cgcaagatcc actacatgaa cacacgcaag cggggtatca acacattagt gagcattatg
     3361 tccagatttg gcaaggagca tgagactgtt tcgaacataa cccacaatga gagactagag
     3421 ttcctcggtg atgctgtagt agaattcctc agctcaatcc atctgttttt tatgttccct
     3481 gaactggagg agggtggttt ggccacttac cgagcggcaa ttgtccaaaa ccagcacttg
     3541 gctctgttgg ccaaaaagct gcaactggaa gagtttatgc tgtacgcaca cggatccgat
     3601 ctgtgccacg agttggaact gcgtcacgcc atggccaact gttttgaagc cctaatgggc
     3661 gcgcttctat tggatggtgg aatcaaggtg gcagatgagg tgttcacgga tgcactcttc
     3721 cgacaggacg agaaactgct gagtatatgg aagaatctgc cggagcaccc gttgcaggaa
     3781 caggagccac ttggggatcg cagctgtatt gattcctacc gtgtgcttaa ggagttaacc
     3841 aaattcgagg actctattgg aatcaagttt aagcacattc ggcttttggc tcgcgctttt
     3901 accgatcgct ccattggttt cacccacttg acccttgggt caaatcagcg tttagagttc
     3961 ttgggcgaca cagtgctgca gctgatttgc tcggagtacc tatatcgtca cttccctgag
     4021 caccacgagg gccacttgtc cctgctacga tcctcattgg tcaataatcg cactcaagcg
     4081 gtggtttgcg atgatttggg aatgccgaaa tatgccgtgt atgccaatcc caaggctgat
     4141 ttgaagacca aagatcgtgc cgatctgctg gaggcattcc tcggcgccct gtacgtggac
     4201 aagggtctcc tgtattgtga acagttttgc catgtatgtt tgttcccgcg acttcagttg
     4261 tttatcatga atcaggactg gaacgatccc aagtcgaagc tgcagcagtg ctgcctcaca
     4321 cttcgcacaa tggatggcgg cgagccggac attccctact acaaggtggt ggaggccagt
     4381 ggtccaacta atacgcgggt atacaaggtg gccgtttatt tccgctcaaa gcggctggcc
     4441 acttcaagtg gctcctccat tcagcaggcg gagatgaacg ccgctaagca ggcgctggag
     4501 aattcaaggg acctgtttcc ccaactggat caccaaaagc gcgtgatcgc caagagcata
     4561 aagaaacaaa cgggcaacga attggacaac gacagtgatc gccagcatca agaagaaaaa
     4621 atcaagcggc ccaaatatgc gacaccgctt caggatgaga gccatcttcc taagcaatat
     4681 cgcatgcacg agaacatttc cagcgatgag ttgccggaag acgaagactt cgaaagcact
     4741 gctcccaaga gtccaacaat gcccttaaga agtaacagaa gtggcagtag tagcagcagc
     4801 agcagcagtg atagcgatgg ctcaacctct tctccgaagt gcaagcgccg attaaagaaa
     4861 tgttcctcgg tgtcttcaaa atcttcgctg ggatgatcta gagcaagggc gaattcgcgg
     4921 ccgcgaattc atcgatagat ctgatatcgg taccagtcga ctctagagga tcccgggtgg
     4981 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     5041 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     5101 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     5161 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     5221 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     5281 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     5341 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     5401 cccaaattgc tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgatttta
     5461 aaataactat accagcagga ggacgtccag acacagcata ggctacctgg ccatgcccaa
     5521 ccggtgggac atttgagttg cttgcttggc actgtcctct catgcgttgg gtccactcag
     5581 tagatgcctg ttgaattggg tacgcggcca gcttggctgt ggaatgtgtg tcagttaggg
     5641 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     5701 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5761 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     5821 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     5881 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     5941 ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag
     6001 tgagtcgtat taaattcgta atcatgtcat agctgtttcc tgtgtgaaat tgttatccgc
     6061 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6121 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6181 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6241 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6301 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6361 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6421 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6481 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6541 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6601 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6661 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6721 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6781 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6841 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6901 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6961 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     7021 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     7081 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7141 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7201 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7261 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7321 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7381 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7441 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7501 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7561 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7621 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7681 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7741 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7801 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7861 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7921 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7981 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     8041 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     8101 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8161 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     8221 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     8281 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     8341 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     8401 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     8461 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     8521 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     8581 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc
     8641 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8701 ccagctggcg aaagggggat g
//
