LOCUS       sequence-212172-        5762 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged D. melanogaster Hel25E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5762)
  AUTHORS   Huang C, Liang D, Tatomer DC, Wilusz JE
  TITLE     A length-dependent evolutionarily conserved pathway controls nuclear
            export of circular RNAs.
  JOURNAL   Genes Dev. 2018 May 17. pii: gad.314856.118. doi: 
            10.1101/gad.314856.118.
  PUBMED    29773557
REFERENCE   2  (bases 1 to 5762)
  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: "Genes Dev. 
            2018 May 17. pii: gad.314856.118. doi: 10.1101/gad.314856.118."
COMMENT     Sequence Label: Hy_pMT Flag-Hel25E WT
FEATURES             Location/Qualifiers
     source          1..5762
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     912..931
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(951..1028)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1234..1253
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1337..1356
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1337..1355
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1356..1380
                     /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)."
     RBS             1395..1417
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     1430..1492
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             1511..1528
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1599..1617)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1613..1660
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1835..1840,1841..1857,1858..1863))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1890..1908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1976..2080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2081..2149,2150..2941)
                     /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     complement(2299..2318)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3112..3700
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3601..3620
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3854..3871
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3886..4025
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3951..3970)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4111..4133
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4127..4318)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5630..5762,1..950))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
       61 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
      121 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
      181 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
      241 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
      301 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
      361 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
      421 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
      481 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
      541 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
      601 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
      661 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
      721 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
      781 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
      841 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
      901 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
      961 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     1021 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     1081 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     1141 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     1201 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     1261 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat
     1321 ctcgatcccg cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc
     1381 cctctagaaa taattttgtt taactttaag aaggagatat accatgggaa tgaaaaagac
     1441 agctatcgcg attgcagtgg cactggctgg tttcgctacc gtagcgcagg ccgctccgaa
     1501 agataacacc caccaccacc accaccacgc ggccgcgggt accctcgagg atccggctgc
     1561 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1621 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     1681 cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
     1741 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
     1801 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
     1861 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     1921 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     1981 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2041 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2101 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2161 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2221 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2281 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     2341 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2401 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2461 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     2521 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2581 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     2641 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2701 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2761 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2821 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2881 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2941 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3001 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3061 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3121 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3181 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3241 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3301 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3361 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     3421 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     3481 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     3541 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     3601 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     3661 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     3721 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     3781 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     3841 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     3901 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     3961 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     4021 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4081 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4141 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     4201 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     4261 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     4321 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     4381 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     4441 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     4501 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     4561 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     4621 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     4681 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     4741 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     4801 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     4861 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     4921 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     4981 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     5041 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat
     5101 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     5161 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     5221 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc
     5281 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     5341 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     5401 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     5461 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     5521 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     5581 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     5641 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5701 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     5761 gc
//
