LOCUS       sequence-197673-        5415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mTagRFPT-linker sequence for N-terminus tagging of
            human CETN2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5415)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5415)
  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            "Unpublished"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: AICSDP-22:CETN2-mTagRFP-T
FEATURES             Location/Qualifiers
     source          1..5415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..75
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     309..326
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    443..464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(479..484,485..502,503..509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    517..533
                     /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     522..544
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     541..557
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     541..557
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(1607..2335)
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(3356..3373)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3356..3372)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3365..3387)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3581..3600)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3700..3722
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3760..3778)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3846..3950
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3951..4019,4020..4811)
                     /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(4169..4188)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4982..5415,1..155)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
       61 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      121 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      181 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      241 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      301 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      361 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      421 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      481 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      541 caggaaacag ctatgaccat gattacgcca agctcggcgc gccattggga tcaggtcacc
      601 ctttcctttt taacaatgta actcgagtct agacttgcgg caagtgtcag gaagcccatt
      661 ttggcctcaa agaaacacca ggcaagccag gcactccatc caaggtgacg tttgctacct
      721 atcgctatat ctctccctat cccatatcta tttcctaact gatttggaat tctgtgggaa
      781 gaggtatctg cttggaagtc ttgagaaaga agcacaaggg cttgttttgg gagcccaaag
      841 taacatgatg ggagggcagg cctcctgatg cagttgctac ctggactgct ctgtggtcac
      901 atgactgttt taacaggaag acgccatgga cctctgcaga tgactggggc tccataccat
      961 gataattgtc ttagctactt accattttct gggtcatcac agttaaaaag tcaccaaagt
     1021 tcatttttcc tgtcccttcc ttatcaattt cacttatcat tttcttaatt tcttctttct
     1081 tgggttcaaa gcccagggcc ctcattgcca cctataaaga aaacaggatc gtctcaataa
     1141 gcacatctaa agtagcagga gcaaaaataa cagtgttgag agcattaagc cagcatctga
     1201 tgtggtcatg ttaatatgat tggggaacac tatgacattg atctaattga tatttgcaat
     1261 caattttatt gaataatact caaagtaagt aaagacagga aaacagcaca gtacaacctc
     1321 ccctgttact tactgcaggg agaagagggc aaaaggggta gagaggtgat agggagatga
     1381 caaggaagag gcagaggaaa ggaagggcag tacggcagga atgcacagag cttgccttca
     1441 gttctttaac atctatggtg ccagttccat ccgcatcgaa aagatcaaaa gcttcccgga
     1501 tctcctgctt ttgctcttca gtaagctcag gcttaggact cattcttttt cgctgagaac
     1561 ttgatgccat gtttgccttt ttgaaattgg agcttctcag gccgctcttg tacagctcgt
     1621 ccatgccatt aagtttgtgc cccagtttgc tagggaggtc gcagtatctg gccacagcca
     1681 cctcgtgctg ctcgacgtag gtctctttgt cggcctcctt gattctttcc agtctgtggt
     1741 ccacatagta gacgccgggc atcttgaggt tcttagcggg tttcttggat ctgtatgtgg
     1801 tcttgaagtt gcagatcagg tggcccccgc ccacgagctt cagggccatg tcggttctgc
     1861 cttccaggcc gccgtcagcg gggtacagca tctcggtgtt ggcctcccag ccgagtgttt
     1921 tcttctgcat cacagggccg ttggatggga agttcacccc tctgatcttg acgttgtaga
     1981 tgaggcagcc gtcctggagg ctggtgtcct gggtagcggt cagcacgccc ccgtcttcgt
     2041 atgtggtgac tctctcccat gtgaagccct cagggaagga ctgcttaaag aaatcgggga
     2101 tgccctgggt gtggttgatg aaggttctgc tgccgtacat gaagctggta gccaggatgt
     2161 cgaaggcgaa ggggagaggg ccgccctcga ccaccttgat tctcatggtc tgggtgccct
     2221 cgtagggctt gccttcgccc tcggatgtgc acttgaagtg gtggttgttc acggtgccct
     2281 ccatgtacag cttcatgtgc atgttctcct taatcagctc ttcgccctta gacacggcct
     2341 ttatatgtta tgcaatacac aagcacaata atatatcata cagtccatat ttacataaca
     2401 tagttacaga cacccatatg tatttttaaa gtcatttata taaccaattc caaagctttg
     2461 cttaaaaaga aagataacag ggtttactgg caagaacaat gaatctttcc atgaacttct
     2521 acccctccct tttctttact gttgtttcta gaagcaggga gatcacttgt tcagtaagga
     2581 cacctcttac tgaaaatcat cccaaaagag agccagtcaa gaagatgact gtgaataact
     2641 agagataatc agctcttccc gaacgtgggg atcctggttc tgaaaacagt cttacgatgc
     2701 ccactcccag tactcccctt cccagtaagt actccgactt cccaattact tctagatttg
     2761 gccaaatatt agtaagttgt taagatgtat attgtgtcag taccgagtaa taaaactcaa
     2821 tccatctggc accttcgtgc cttctccaag aagctaaagc atcccctgaa tgcccctgta
     2881 tttaaattgt aaaaaacatt gtaacagaac acgatacacc ttttcccagc tgtgtgtgga
     2941 aattttggca gaatgaggag gccagggggc caaggtctga gagagtcagg ggcctgtgag
     3001 ggattcgagt cttttttgcc cagggaggtt caccgagggg atcagggaat gcaaggggag
     3061 cccgaggggc gggtgtgggc ctgttgtggg tgaagaggag gcagcagcga agcgcaggcc
     3121 cagtaggttg gttagcgtct gtaccctact ctcccgggaa agagctcaaa ggaaagactg
     3181 gtgcgctaaa gccgagaaag gcgcaggaga ggcggcgtgg ccgagctggg gcctgggaag
     3241 ggctcagccg ctgggcgtgg ggcgcgacag agcggcagca ctcaccatag ccaaaggagt
     3301 ccgctgccgg ttgttaggca accgacgtgt acactatccc aatacgcgtc aattcactgg
     3361 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3421 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3481 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     3541 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     3601 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3661 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3721 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3781 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3841 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3901 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3961 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4021 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4081 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4141 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4201 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4261 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4321 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4381 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4441 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4501 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4561 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4621 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4681 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4741 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4801 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4861 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4921 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4981 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5041 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5101 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5161 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5221 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5281 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5341 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5401 ggagaaaggc ggaca
//
