LOCUS       sequence-342165-        6636 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express 3xFLAG-Kctd19 under the mouse Clgn promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6636)
  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 6636)
  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_3xFLAG-Kctd19
FEATURES             Location/Qualifiers
     source          1..6636
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     238..258
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        240..258
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      618..627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             627..692
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3586..3605)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3651..3706
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3705..3724)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4061..4080)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4062..4080)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4090..4107)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4090..4106)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4099..4121)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4248..4703
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4335..4354)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4545..4566
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4729..4833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4834..4902,4903..5694)
                     /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(5052..5071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5865..6453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6354..6373
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6607..6624
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca
      241 attaaccctc actaaaggga acaaaagctg gagctcatcg atgagctcca gagagggact
      301 gacgttggaa ggaccgcatc cccccaggcg cgcggccgct gggtcccgag cggcggttga
      361 gcgggccgct tgcgcactgg ggctgccact gcatcctgca tcctccggcg cggccgctcg
      421 cgcacgcgca gttagcagtc gctgctgggc ggctgccggc gggatcgctc tggggagacg
      481 ccgggcacca agtagcggcc agtcggacac ctgagctgcc gctgctggac acaagcccgc
      541 aggacaggtg tgtagaccgc gggccgcctt cctgcggctt gttctctggg gaccgggtga
      601 gaaatcggat ctctagagcc gccatggact acaaagacca tgacggtgat tataaagatc
      661 atgacattga ttacaaggat gacgatgaca aggggggcgg ggctagcgga tctgaggagc
      721 ctggcgggct ccatgagtca gcagaggatt tgtttcactt caatgtcggg ggctggcatt
      781 tctcagttcc caggagcaaa cttgctcagt tcccagactc gctgttgtgg aaagaggctt
      841 cagccctgac ctcttcagag aaccagcggc tcttcattga cagagacggc tccacgttta
      901 ggcatgtgca ctattacctc tacacgtcca agctctcctt ctccagttgt gcagaactga
      961 acttgctcta tgagcaagcc ctgggtttgc agctgatgcc cttgcttcag accctagata
     1021 acctgaagga agggagacac cacctccgtg tgcggcctgc cgacattcct gtggctgaga
     1081 gagcatccct gaactactgg cggacatgga agtgtatcag caagccctcg gattttccaa
     1141 taaaaagccc cgccttcaca ggtcttcatg acaaggcccc tttgggactc atggacacac
     1201 cgctgttgga cacagaagag gaagtgcact actgcttctt acccctcgac ttggtggcca
     1261 agtacccaag cctagtgacc gaagacaatc tgctgtggct ggctgaaacg gtggccctga
     1321 tcgagtgcga atgcagcgag ttccgcttca ttgtgaattt tctacgctca cacaagattt
     1381 tactaccaga taacttctcc aacatagatg tgttagaagc agaagtagaa attctggaaa
     1441 tccctgagct tacagaagct gtacggttgt accggatgaa tatgggtggc tgttcccgga
     1501 cctcctgccc acctctaagt cctggaaagg ggggtcgtac agccagtgtg gagtctgtga
     1561 aaccactgta cttgatggcc ctgggcctgc tggtcaagta tccagactct gcactgggac
     1621 agctgcgcat cgagagcacg ctggacggaa gcagactata catcacaggg aacggggccc
     1681 ttttccagca cgtcaggaac tggctgggca cttgtcggct gcccctgact gagaccatct
     1741 ccgaggtgta tgagctctgc gccttcctgg acaagaggga catcacctac gagccaatga
     1801 aagtggctct gaagactcat ctggagccaa ggactttgct gcccgtggat gtgctcagtg
     1861 aggagtggac agctgaggtc acgatatact caccacagca gatcatcaag ctttatgttg
     1921 gaagccattg gtatgcaacg accctgcaga ccctgatgaa gtacccagaa ctgctatcca
     1981 acactcagag agtgtactgg atcgcctacg gacagacctt gctcattcac ggggacggcc
     2041 aaatgttccg acacattctc aacttcctga gacttggaaa actgttttta ccatcagaat
     2101 ttaaggaatg gcctctcttc tgccaggagg tggaggagta tcatattcca gccctctcag
     2161 aagccctggc acaatgcgaa gcatacaagt catggactca ggagaaagaa tctgaaaatg
     2221 aagaagcttt tcccattagg aagctgcatg tggtgaccga agggacaggg ccaatggcag
     2281 agttcagcag agatgccaag gaaaccacag cctgcatgcc tgtggacttc caggaatgca
     2341 gtgacaggac tccatggaac aaggccaaag ggaacctgac caggtccagc cagatggaag
     2401 aggctgagca gtatacccgg actattcagg tctccctctg cagaaatgcc aagaggggag
     2461 gcaatcccag cacatactca cactgctctg gtttatgtgc caaccccaga cattggggag
     2521 gtcactctga gagtcccccc aagaagaagt gcaccactat aaacctcaca cagaaacctg
     2581 atgccaaaga ccctcctgtc acccccatgc aaaaactcat atctttagtc agggaatggg
     2641 acatggtcaa ctgtaaacag tgggaattcc agccactgcc agcctccagg agcagctctg
     2701 tggaggaggc tagcctgcat gtcccctcgg gaagtgaggc tgctccgcag cctggtacct
     2761 cagctgcttg gaaagctcat tccataacct ctgagaagga tgctggtccc cagactggac
     2821 ccggagctgg agtgaaagac aagggtccag agccaacctt taagccatac ttccctacaa
     2881 aaagagctat taccctaaag gactggggca agcagaggcc caaggacaga gaaagccctg
     2941 ctcctgagca gcctctgcct aatgctaacg ggaccgacaa tccaggggcc atcctcaaag
     3001 tggctcaccc ccctgtggtg ggtaatgatg gctcctgcat gttctttgaa gacagcatta
     3061 tctacaccac ccaaatggat aacatcaagc atacatcact cacagccagc ccccaacttc
     3121 gagaagtgac ttttctgagc ttctctctgt cctgggagga gatgttttat gcacagaaat
     3181 gtcaccgctt cctgaccgac atcatcttgg attccatcag gcaaaaggat cccaaagcta
     3241 tcacagccaa ggtggtgtct ctggcctacc gactgtggac actgaacatc agccccaagc
     3301 agttcgtggt ggatctgctg gccatcgctg gcttcaagga tgaccggcac actcaggagc
     3361 gtctgtacag ttgggtagag ctcacactgc ctttcgccag aaaatatggc cgttgcgtgg
     3421 acctgctcat ccagagggga ctgtccaggt ctgtctctta ttcagtcctg ggcaagtatt
     3481 tacacgaggg ctaggcggcc gcctcgagac tcctcaggtg caggctgcct atcagaaggt
     3541 ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc
     3601 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     3661 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3721 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     3781 tgcccatatg ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa
     3841 acagccccct gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt
     3901 ttttatattt tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt
     3961 tttactagcc agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct
     4021 cttatgaaga tccctcgact taattaaggt acccaattcg ccctatagtg agtcgtatta
     4081 cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     4141 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     4201 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag
     4261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     4441 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     4501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     4561 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     4621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     4681 caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct
     4741 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4801 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4861 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4921 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4981 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5041 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5101 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5161 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5221 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5281 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5341 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5401 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5461 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5521 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5581 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5641 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5701 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5761 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5821 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5881 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5941 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6001 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6061 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6121 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6181 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6241 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6301 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6361 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6421 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6481 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6541 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6601 gagcgcagcg agtcagtgag cgaggaagcg gaagag
//
