LOCUS       sequence-79729-e        7207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression in Hydra for RNAi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  AUTHORS   Juliano CE, Reich A, Liu N, Gotzfried J, Zhong M, Uman S, Reenan RA,
            Wessel GM, Steele RE, Lin H
  TITLE     PIWI proteins and PIWI-interacting RNAs function in Hydra somatic 
            stem cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Jan 7;111(1):337-42. doi: 
            10.1073/pnas.1320965111. Epub 2013 Dec 23.
  PUBMED    24367095
REFERENCE   2  (bases 1 to 7207)
  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"; doi: 
            "10.1073/pnas.1320965111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-01-7- 7"; volume: "111"; issue: "1"; pages: "337-42"
COMMENT     Sequence Label: pHyVec12
FEATURES             Location/Qualifiers
     source          1..7207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5019..5039)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5019..5037)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5058..5074)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5058..5074)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5071..5093)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5082..5098
                     /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(5106..5112,5113..5130,5131..5136))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5151..5172
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5289..5306)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5460..6048)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5540..5559)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6219..7010,7011..7079))
                     /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     6842..6861
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7080..7184)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggttttaag
      661 ccgagatgca ggattctgaa tgaaaaagaa aaaaagaagt ctcggtagag taaaagtgat
      721 cggtttgcaa ctgtaaaatt tattgaagta ccaataattt tatttaaaat aaaactgaaa
      781 tataaagtta aagttgctgt tctataagtt tactcgaatt ttaaaaccat tgtaacgcta
      841 gagtaatatt tgagtctact aagttagtcc ccgcactttt taatcaagca ataaataccc
      901 aaactttgct tattcaaatc aataaaccaa tatatctctt aaaataaagt aaaaacttct
      961 gaaattctat aaaaaaaaat ttaatttcga aatatcaaat gtaacttcaa caccgcacta
     1021 ttttctttta aacaactgat atagtaatta cttctcaaaa acgttatctc aaggtttgtg
     1081 atgtacttaa aaccactcct attttgttac gcgtttaaaa aagcaaacat aagttggttt
     1141 ctattgatga atgagaacat atttcattta aagttaaaat cctaccagtg gtttcactgt
     1201 acgtaaacac cgtcaaaaaa acaggaacgt ttttaaagat taataattga agtaaaaaaa
     1261 atttaatacc gggggttaaa aaaatctttt aaaataatta taaatatata tattaaaatt
     1321 tataaatttt taaacacatt taaaatatat attaagtata ataaaagtaa tattataaaa
     1381 aaaaatttaa ttttataatt atttttatta aatttataaa taataggtaa aacttacata
     1441 tccgttttat tttttcttaa taaaataacg cgtgcaaatt tttgtccata taaagacctt
     1501 ttcgaacaat aacttttttg cttagccgtt ttttttctta tatggtcaaa aaagcgctca
     1561 agcgattcac cataaaaagc gcaattagtt cagcgttcgt tattcagaag cttcagcttt
     1621 gcttgataat cagctcttct ctttttaaac aaaacactta atcaagctag caaaaatgag
     1681 taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
     1741 taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact
     1801 tacccttaaa tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac
     1861 tactttctgt tatggtgttc aatgcttttc aagataccca gatcatatga aacggcatga
     1921 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga
     1981 tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag
     2041 aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga
     2101 atacaactat aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa
     2161 agttaacttc aaaattagac acaacattga agatggaagc gttcaactag cagaccatta
     2221 tcaacaactg caggtaacta actgggatga tatggaaaaa gtaagactta ttattattaa
     2281 attaaaactt agctttattt aaatcgaaat ttagctagta tatcagtata atttaattca
     2341 aaattactta gtcattaatt tcgagtcctt aacaacttca atttatttta attaaattat
     2401 aaagtagcaa tcattggcca aattatggcg aactttatca gcaaaattag tgtaaaatca
     2461 aaatcatttt tgtgaaatat ctaaataagt agcaaataac aaattagtat taccaataaa
     2521 taatgtaaat taactatgat taattaagtt taaagtagct ccataaagac taatatttta
     2581 atatctataa taacattttg caaactttaa atttttttat tattattaat tttatacttt
     2641 gttaataaaa taatcaaaaa tataaaccag atcataaatt gtatacatat tttacttatc
     2701 aattttattt gtaaattata ataatttttc atttatcatt tagatttggc atcacacttt
     2761 ttacaatgag ctccgtgttg ctctcgagtt gttggataat ggtctgctag ttgaacgctt
     2821 ccatcttcaa tgttgtgtct aattttgaag ttaactttga ttccattctt ttgtttgtct
     2881 gccatgatgt atacattgtg tgagttatag ttgtattcca atttgtgtcc aagaatgttt
     2941 ccatcttctt taaaatcaat accttttaac tcgattctat taacaagggt atcaccttca
     3001 aacttgactt cagcacgtgt cttgtagttc ccgtcatctt tgaaaaatat agttctttcc
     3061 tgtacataac cttcgggcat ggcactcttg aaaaagtcat gccgtttcat atgatctggg
     3121 tatcttgaaa agcattgaac accataacag aaagtagtga caagtgttgg ccatggaaca
     3181 ggtagttttc cagtagtgca aataaattta agggtaagtt ttccgtatgt tgcatcacct
     3241 tcaccctctc cactgacaga aaatttgtgc ccattaacat caccatctaa ttcaacaaga
     3301 attgggacaa ctccagtgaa aagttcttct cctttactca tttttgataa cgaggatcca
     3361 taaatctgat tgcgatcttt tgttttccaa taaaacaaag ataccgaaag ttaccgattt
     3421 ttcttaccga aagtttgaca atgctttgtt gttctccatt tttctattgt aattaaaaac
     3481 attttttctt tttttaagaa gttgtgcttt ttataaattt gtgtaaaaac gcgttataat
     3541 gctaagttga gggttaaagt ttactgacct gataatatat cgagcgagga ttattatata
     3601 attaaagatt tattagtata tatagtatat aacatgtaaa ctgatagtat atacttgata
     3661 tattaatgat taatatatca agcgaggatt catttatcat taaagtattt gcgatcgtcg
     3721 cttgtaacat taaattttaa attttacctt attaaaactt cataattttt tttattttta
     3781 gatagtaaat atagtagcaa tggcagatgc tgcagaaatg gcttcatcag aaaatgttat
     3841 tacagaattt atgagattta aagttagaat ggaaggtaca gttaatggac atgaatttga
     3901 aattgaggga gaaggagaag gtagaccata tgaaggacat aatacagtta aattaaaggt
     3961 tacaaaagga ggaccattac catttgcgtg ggatatttta tcaccacaat ttcaatatgg
     4021 atcaaaggtt tacgttaaac atccagctga tattccagat tataaaaaat tatcttttcc
     4081 agaaggattt aaatgggaaa gagttatgaa ttttgaagat ggtggagttg ctacagttac
     4141 acaaagattt catcattcca agatggttgt tttatttata aagttaaatt tattggggtt
     4201 aattttccat cagacggacc agttatgcaa aaaaaaacaa tgggatggga agctagtaca
     4261 gaaagattat atccaagaga cggggtttta aagggagaaa cacataaagc tctaaaatta
     4321 aaggacggag gacattattt agttgaattt aaatcaattt atatggctaa aaaaccagtt
     4381 caattaccag gatattatta tgtagatgct aagttagata ttacatcaca taatgaagat
     4441 tatacaattg ttgaacaata tgaaagaaca gaaggtagac atcatttatt tttatagcat
     4501 tcgtagattc caagggcgaa ttcacaattc gattatattt atactggact atttttacat
     4561 ctgttcggtt attttcacat ttatttttct atatatatct tataaacgtt ttaaaaccca
     4621 tgtaattttt gttaagctgt aatataaaag acgtcctaac aaacttcttt tattactgaa
     4681 tttcctttaa ttataataaa taacaagttt taaaataaat tcaggcaatt aaggcgctcc
     4741 tgaggtacta aaattaatgt aaacatttaa aattaacttg gatggtctta agtactgtac
     4801 tcgtgatttt gttatacttt attattagaa aagtcgtcta ttaacttttt gttccttaat
     4861 ttacttgatt aaattgtcgc ttaatttatc aaatcaggtt ttgcgcgtta ttttagagaa
     4921 aaacttatta gaaaaatgaa taagcaaagt ttaggctaac atgttttttt attattttaa
     4981 atagttcaag tcaatgacgt ataaaccagc ttttgttccc tttagtgagg gttaattgcg
     5041 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     5281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5641 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5881 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     7021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     7081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     7141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     7201 gtgccac
//
