LOCUS       sequence-341875-        6685 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express Kctd19-3xHA under the mouse Clgn promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6685)
  AUTHORS   Oura S, Koyano T, Kodera C, Horisawa-Takada Y, Matsuyama M, Ishiguro
            KI, Ikawa M
  TITLE     KCTD19 and its associated protein ZFP541 are independently essential
            for meiosis in male mice.
  JOURNAL   PLoS Genet. 2021 May 7;17(5):e1009412. doi: 
            10.1371/journal.pgen.1009412. eCollection 2021 May.
  PUBMED    33961623
REFERENCE   2  (bases 1 to 6685)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Genet."; date: "2021-05-7"; volume: "17(5):e1009412. doi"; pages: " 
            10.1371/journal.pgen.1009412. eCollection 2021 May"
COMMENT     Sequence Label: pClgn1.1_Kctd19-3xHA
FEATURES             Location/Qualifiers
     source          1..6685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(20..25,26..43,44..50)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    58..74
                     /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     63..85
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     82..98
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     82..98
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     117..137
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        119..137
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      533..542
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3338..3427
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(3514..3533)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3579..3634
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3633..3652)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3989..4008)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3990..4008)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4018..4035)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4018..4034)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4027..4049)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4176..4631
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4263..4282)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4473..4494
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4657..4761
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4762..4830,4831..5622)
                     /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(4980..4999)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5793..6381
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6282..6301
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6535..6552
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    join(6669..6685,1..5)
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
       61 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa
      121 ttaaccctca ctaaagggaa caaaagctgg agctcatcga tgagctccag agagggactg
      181 acgttggaag gaccgcatcc ccccaggcgc gcggccgctg ggtcccgagc ggcggttgag
      241 cgggccgctt gcgcactggg gctgccactg catcctgcat cctccggcgc ggccgctcgc
      301 gcacgcgcag ttagcagtcg ctgctgggcg gctgccggcg ggatcgctct ggggagacgc
      361 cgggcaccaa gtagcggcca gtcggacacc tgagctgccg ctgctggaca caagcccgca
      421 ggacaggtgt gtagaccgcg ggccgccttc ctgcggcttg ttctctgggg accgggtgag
      481 aaatcggatc tctagaggat ccctgcagga attcgatatc aagcttgtcg acgccgccat
      541 ggaggagcct ggcgggctcc atgagtcagc agaggatttg tttcacttca atgtcggggg
      601 ctggcatttc tcagttccca ggagcaaact tgctcagttc ccagactcgc tgttgtggaa
      661 agaggcttca gccctgacct cttcagagaa ccagcggctc ttcattgaca gagacggctc
      721 cacgtttagg catgtgcact attacctcta cacgtccaag ctctccttct ccagttgtgc
      781 agaactgaac ttgctctatg agcaagccct gggtttgcag ctgatgccct tgcttcagac
      841 cctagataac ctgaaggaag ggagacacca cctccgtgtg cggcctgccg acattcctgt
      901 ggctgagaga gcatccctga actactggcg gacatggaag tgtatcagca agccctcgga
      961 ttttccaata aaaagccccg ccttcacagg tcttcatgac aaggcccctt tgggactcat
     1021 ggacacaccg ctgttggaca cagaagagga agtgcactac tgcttcttac ccctcgactt
     1081 ggtggccaag tacccaagcc tagtgaccga agacaatctg ctgtggctgg ctgaaacggt
     1141 ggccctgatc gagtgcgaat gcagcgagtt ccgcttcatt gtgaattttc tacgctcaca
     1201 caagatttta ctaccagata acttctccaa catagatgtg ttagaagcag aagtagaaat
     1261 tctggaaatc cctgagctta cagaagctgt acggttgtac cggatgaata tgggtggctg
     1321 ttcccggacc tcctgcccac ctctaagtcc tggaaagggg ggtcgtacag ccagtgtgga
     1381 gtctgtgaaa ccactgtact tgatggccct gggcctgctg gtcaagtatc cagactctgc
     1441 actgggacag ctgcgcatcg agagcacgct ggacggaagc agactataca tcacagggaa
     1501 cggggccctt ttccagcacg tcaggaactg gctgggcact tgtcggctgc ccctgactga
     1561 gaccatctcc gaggtgtatg agctctgcgc cttcctggac aagagggaca tcacctacga
     1621 gccaatgaaa gtggctctga agactcatct ggagccaagg actttgctgc ccgtggatgt
     1681 gctcagtgag gagtggacag ctgaggtcac gatatactca ccacagcaga tcatcaagct
     1741 ttatgttgga agccattggt atgcaacgac cctgcagacc ctgatgaagt acccagaact
     1801 gctatccaac actcagagag tgtactggat cgcctacgga cagaccttgc tcattcacgg
     1861 ggacggccaa atgttccgac acattctcaa cttcctgaga cttggaaaac tgtttttacc
     1921 atcagaattt aaggaatggc ctctcttctg ccaggaggtg gaggagtatc atattccagc
     1981 cctctcagaa gccctggcac aatgcgaagc atacaagtca tggactcagg agaaagaatc
     2041 tgaaaatgaa gaagcttttc ccattaggaa gctgcatgtg gtgaccgaag ggacagggcc
     2101 aatggcagag ttcagcagag atgccaagga aaccacagcc tgcatgcctg tggacttcca
     2161 ggaatgcagt gacaggactc catggaacaa ggccaaaggg aacctgacca ggtccagcca
     2221 gatggaagag gctgagcagt atacccggac tattcaggtc tccctctgca gaaatgccaa
     2281 gaggggaggc aatcccagca catactcaca ctgctctggt ttatgtgcca accccagaca
     2341 ttggggaggt cactctgaga gtccccccaa gaagaagtgc accactataa acctcacaca
     2401 gaaacctgat gccaaagacc ctcctgtcac ccccatgcaa aaactcatat ctttagtcag
     2461 ggaatgggac atggtcaact gtaaacagtg ggaattccag ccactgccag cctccaggag
     2521 cagctctgtg gaggaggcta gcctgcatgt cccctcggga agtgaggctg ctccgcagcc
     2581 tggtacctca gctgcttgga aagctcattc cataacctct gagaaggatg ctggtcccca
     2641 gactggaccc ggagctggag tgaaagacaa gggtccagag ccaaccttta agccatactt
     2701 ccctacaaaa agagctatta ccctaaagga ctggggcaag cagaggccca aggacagaga
     2761 aagccctgct cctgagcagc ctctgcctaa tgctaacggg accgacaatc caggggccat
     2821 cctcaaagtg gctcaccccc ctgtggtggg taatgatggc tcctgcatgt tctttgaaga
     2881 cagcattatc tacaccaccc aaatggataa catcaagcat acatcactca cagccagccc
     2941 ccaacttcga gaagtgactt ttctgagctt ctctctgtcc tgggaggaga tgttttatgc
     3001 acagaaatgt caccgcttcc tgaccgacat catcttggat tccatcaggc aaaaggatcc
     3061 caaagctatc acagccaagg tggtgtctct ggcctaccga ctgtggacac tgaacatcag
     3121 ccccaagcag ttcgtggtgg atctgctggc catcgctggc ttcaaggatg accggcacac
     3181 tcaggagcgt ctgtacagtt gggtagagct cacactgcct ttcgccagaa aatatggccg
     3241 ttgcgtggac ctgctcatcc agaggggact gtccaggtct gtctcttatt cagtcctggg
     3301 caagtattta cacgagggcg atatcggggg cgggggctac ccatacgatg ttcctgacta
     3361 tgcgggctat ccctatgacg tcccggacta tgcaggatcc tatccatatg acgttccaga
     3421 ttacgcttag ctcgagactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     3481 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     3541 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3601 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3661 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3721 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     3781 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     3841 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     3901 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc
     3961 cctcgactta attaaggtac ccaattcgcc ctatagtgag tcgtattacg cgcgctcact
     4021 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     4081 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     4141 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     4201 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     4261 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4321 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4381 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4441 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4501 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     4561 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4621 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4681 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4741 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4801 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4861 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4921 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4981 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5041 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5101 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5161 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5221 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5281 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5341 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5401 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5461 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     5521 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5581 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     5641 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5701 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5761 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5821 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5881 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     5941 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6001 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6061 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6121 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6181 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6241 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6301 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6361 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6421 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     6481 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     6541 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     6601 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     6661 aacgcaatta atgtgagtta gctca
//
