LOCUS       sequence-291833-        8049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Flag-tagged hRPGRIP1L in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8049)
  AUTHORS   Takemaru et al.
  TITLE     Takemaru lab unpublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8049)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Flag-hRPGRIP1L
FEATURES             Location/Qualifiers
     source          1..8049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1069..1092
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5079..5213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5103..5122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5157..5176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5326..5346)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5326..5344)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5365..5381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5365..5381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5378..5400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5389..5405
                     /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(5413..5419,5420..5437,5438..5443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5458..5479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5596..5613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5767..6355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5847..5866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6526..7317,7318..7386))
                     /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     7149..7168
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7387..7491)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(7517..7972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7654..7675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7866..7885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcacagg gccaagaccc ctcctcaaac ttggcaccgt
      181 gccaagtaga cccctcccca gttggcacag tgccaggtcc gccatattga attggcatgg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcaggat ccaccatgga ttacaaggat
     1081 gacgacgata agatctcgaa ttcgtctggt ccaactgatg agactgcagg agacttgcct
     1141 gtgaaagata caggtctaaa cctctttgga atgggagggt tacaagaaac ttcaacaaca
     1201 cggacaatga agtctcgcca ggcagtgtca cgtgtcagtc gtgaggaact ggaagacaga
     1261 tttttgcgtt tgcatgatga gaacatttta cttaaacagc atgcccgcaa gcaggaggat
     1321 aaaattaaaa gaatggccac caagttaata cggctagtta atgacaagaa aagatatgag
     1381 cgggttggtg gcggccccaa gcggctggga cgagatgtgg aaatggaaga aatgattgag
     1441 cagctgcaag agaaagttca tgagcttgaa aaacaaaatg aaaccctcaa aaacagactg
     1501 atttcagcca aacagcaact tcaaacccag ggttacaggc aaactccata caataatgta
     1561 caatctcgta ttaacactgg gcgtagaaaa gcaaatgaaa atgctggttt acaggaatgc
     1621 cccaggaaag gtataaaatt ccaagatgca gatgtagcag aaactccaca tcccatgttt
     1681 acaaaatatg gcaacagttt acttgaagaa gccagaggag aaataagaaa tttagaaaac
     1741 gttattcagt cacaaagagg ccagatagag gagttagagc acttggctga gatcctgaaa
     1801 actcagttga ggagaaaaga aaatgaaatt gagttatctc ttcttcagct tcgagaacag
     1861 caagctacag atcaaaggtc aaatattcgg gacaatgtag aaatgattaa gcttcataaa
     1921 cagctagtag agaaaagcaa tgctctttca gcaatggaag gaaaatttat tcagcttcaa
     1981 gagaagcaaa gaactctcag aatcagccac gatgctttga tggcaaatgg ggatgaatta
     2041 aacatgcaac ttaaagagca gcgtttaaaa tgctgcagtc ttgagaaaca attacattct
     2101 atgaagtttt ctgaaagaag aatagaagag ctgcaggata gaattaatga tttagaaaag
     2161 gaacgggaac ttttaaagga aaactatgat aaactttatg acagtgcctt cagtgctgcc
     2221 catgaagagc aatggaagtt aaaggagcaa cagctgaaag tgcagattgc tcagctcgag
     2281 actgccttga agtctgactt aacagacaaa actgaaatcc ttgacagatt aaaaactgaa
     2341 agagatcaaa atgaaaaact cgttcaagag aatagagaac tacagttaca gtatctggaa
     2401 caaaagcaac aacttgatga acttaaaaaa cgcataaaat tgtacaacca ggagaatgat
     2461 attaatgctg atgaattgag tgaagctctc ctacttataa aggctcaaaa agaacaaaaa
     2521 aatggagacc tttccttttt agtgaaagta gatagtgaaa ttaataaaga tctagaacgc
     2581 tctatgagag agctgcaagc aactcatgca gaaacggtgc aagagctgga aaagacaaga
     2641 aacatgctaa ttatgcaaca caaaattaat aaagattatc agatggaggt tgaggcagtg
     2701 acccgtaaga tggaaaattt gcagcaagat tatgaactca aagtggaaca gtatgttcat
     2761 cttcttgata tcagggctgc acgtatccat aaactagaag cccaattaaa ggatattgcc
     2821 tatgacacca agcagtacaa atttaaacca gaaatcatgc cagatgactc tgttgatgaa
     2881 tttggtgaaa ccatccactt agaacgaggc gaaaatctat ttgaaatcca tatcaacaaa
     2941 gtaacctttt cttctgaagt tttacaggca tctggagata aagagcctgt cactttctgt
     3001 acctatgctt tctatgattt tgaactacag acaactcccg tagtgcgagg ccttcatccc
     3061 gaatataact tcacttctca atatcttgtt catgttaatg acttattttt gcaatatatt
     3121 cagaagaata ctatcaccct tgaggtccac caggcttata gcacagaata tgaaacaatt
     3181 gcagcatgtc aattaaaatt tcacgaaatt cttgaaaaaa gcggccgaat attttgtaca
     3241 gcaagtttga ttggaacaaa aggagacatc ccaaattttg gcacagtgga atactggttc
     3301 cgattaagag ttcccatgga tcaagcaatt cgactttatc gagaaagggc aaaggctttg
     3361 gggtatataa catcaaattt taaggggcca gagcatatgc agtcgttaag tcagcaagca
     3421 cccaaaactg ctcaactcag ttctacagat tccacagatg gcaacttaaa tgaacttcac
     3481 attacaataa gatgttgcaa ccacctgcag tcccgagcaa gccacctgca gccacaccca
     3541 tatgttgtgt acaagttttt tgattttgca gaccatgata cagctatcat tcccagtagc
     3601 aatgatccac agtttgatga tcatatgtat ttcccagtgc caatgaatat ggacttggat
     3661 cgatacctta agtcagagtc tctgagtttt tatgtttttg atgatagtga tacccaggag
     3721 aatatttaca taggaaaagt caatgtgcct ctgatttcgt tggcacatga caggtgtatc
     3781 tcaggaatat ttgagttaac agaccatcaa aagcatcctg ctggcaccat ccatgttata
     3841 ttgaaatgga aatttgctta ccttccacca agtggatcaa taacaactga agacttagga
     3901 aatttcattc gcagcgaaga gccagaagtt gttcaaagac ttcctccagc atcctctgtt
     3961 agcacactag ttttagcacc aagacctaaa ccaagacaac gtttaacacc tgtagataag
     4021 aaggtatctt tcgtggatat catgccacat cagagtgatg agacttctcc tcctcttgaa
     4081 gataggaagg aaatttcacc agaggtagag catataccag aaatagaaat taatatgctg
     4141 actgttccac atgttcccaa ggtttcacaa gaaggcagtg tagatgaggt aaaagagaat
     4201 actgagaaaa tgcagcaagg aaaagatgat gtgtctttac tatctgaagg tcagcttgca
     4261 gaacaaagct tggcatcttc tgaagatgaa acagaaataa cagaggactt ggaaccagaa
     4321 gttgaagagg acatgtcagc ttctgacagt gatgactgta ttattccagg tcctatctcc
     4381 aagaatatca aacagagtct cgcattgtct cccgggctgg ggtgcagtag cgcgatctca
     4441 gctcactgca acttccgcct cccgggttcc agcgattttc ctgcctcagc ctcccaagta
     4501 gatgggatta caggtgcctg ccaccacagc cagccatcag aaaaaattcg gattgagatc
     4561 atagctctaa gccttaatga ttctcaagta accatggatg acactatcca acggctgttt
     4621 gttgagtgtc gattctacag tcttcctgct gaagagacac ccgtgtcact tccaaaaccc
     4681 aagagtgggc agtgggtcta ctataactat agcaatgtga tctacgtgga taaagaaaac
     4741 aacaaagcaa agagagacat cttaaaagct atactacaaa aacaagagat gcctaataga
     4801 agccttcgct tcaccgtggt cagtgaccct ccagaggacg agcaggacct ggagtgtgag
     4861 gacattggcg tggctcacgt cgaccttgcc gacatgtttc aggaagggag ggacctcatt
     4921 gagcaaaata tcgatgtttt tgatgcacga gcagatggtg aaggtattgg caagctcagg
     4981 gtaacagtcg aagctctcca tgccctccag tctgtctaca agcaatacag agatgacttg
     5041 gaggcttgag ctagaactat agtgagtcgt attacgtaga tccagacatg ataagataca
     5101 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     5161 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     5221 acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt ttttaattcg
     5281 cggccgcggc gccaatgcat tgggcccggt acccagcttt tgttcccttt agtgagggtt
     5341 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5401 cacaattcca cacaacatac gagccgggag cataaagtgt aaagcctggg gtgcctaatg
     5461 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5521 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5581 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5641 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5701 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5761 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5821 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5881 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5941 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6001 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6061 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6121 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6181 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6241 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6301 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6361 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6421 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6481 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6541 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6601 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6661 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6721 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6781 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6841 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6901 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6961 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7021 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7081 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7141 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7201 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7261 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7321 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7381 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7441 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7501 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     7561 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     7621 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     7681 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     7741 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     7801 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     7861 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     7921 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     7981 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     8041 cagtcgacc
//
