LOCUS       sequence-197676-        5417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology arms and mTagRFPT-linker sequence for N-terminus (second 
            exon) tagging of human TUBA1B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5417)
  AUTHORS   Allen Institute for Cell Science
  TITLE     Allen Institute for Cell Science Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5417)
  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            "Unpublished"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-28:TUBA1B-mTagRFP-T
FEATURES             Location/Qualifiers
     source          1..5417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(128..716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(208..227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(887..1678,1679..1747))
                     /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     1510..1529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1748..1852)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1920..1938
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1976..1998)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2098..2117
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2311..2333
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2325..2342
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2326..2342
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3363..4091
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(5143..5159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5143..5159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5156..5178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5167..5183
                     /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(5191..5197,5198..5215,5216..5221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5236..5257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5374..5391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
       61 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      121 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      181 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      241 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      301 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      361 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      421 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      481 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      541 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      601 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      661 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      721 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1141 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1201 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1261 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1321 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1381 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1441 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1501 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1561 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1621 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1681 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1741 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1861 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     1921 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     1981 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2041 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     2101 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     2161 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     2221 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     2281 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     2341 gtgaattgac gcgtattggg ataaagcaat aagaggactg cggaagagct ccctgtcaat
     2401 gtaccgctct acaccagtgt attacgacag ttcgtacaca acagtctgta gaggccacct
     2461 gtctctccct gctgcgttag gaattcaggg gagcaggtgg tggcagtaag ggattttgag
     2521 ggaacggaaa tcggatcttg acccagatct gggccgccga taatctccta ctgcgctcag
     2581 actgctgtgg aggtgttagg ctgagcccga tgccggcagg caagggagga tgggcggctt
     2641 gggcagcgcc tttgcagacg tggccatttc gtgcctctgc agcaccgccg gggggcgcaa
     2701 gagcgcgcgc ccggaattgc tcattcatcc tgtgccgcag agccccgccc cttgtccctg
     2761 cggacagaca tttcttctgc gctggtctgg ccacgtgctt cctgtgctag gagctgcccg
     2821 gaaatgtgac cacctagtct aaagtgggct tctggggcct gagcgctgga tggatgccca
     2881 ccttcctgtc ttggtcctcc aaaggaggaa gctgtgactg agctgtcttg gtctggaagg
     2941 aggccttccc ggtttaggat gggaaggtaa cattcattaa aagcaacgta gactatagtg
     3001 tagctgttct caaaagtagt acatcttaga aaaggatctt tagaaaagat cgctttagaa
     3061 aaggaaattc gttttcagat tacgtgagta gcctaggtaa cacagccaga cctcatctcc
     3121 acaaaaaaaa tgaaaaaatt agccagcttg gtggtctgtg cctgtggtcc cagctgctcc
     3181 agaggctgag gtggggggat gactggagcc taggctgcag tgagcctaga tggcatcact
     3241 gcactcaaga ctgggcgaca gaccttatct ctaaaaaaat aaagattgca tgagtatttt
     3301 gttccacttg acagtcatca atagattggt ttaaattgtg atatcttttt tacttaccgc
     3361 aggtgtctaa gggcgaagag ctgattaagg agaacatgca catgaagctg tacatggagg
     3421 gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     3481 gcacccagac catgagaatc aaggtggtcg agggcggccc tctccccttc gccttcgaca
     3541 tcctggctac cagcttcatg tacggcagca gaaccttcat caaccacacc cagggcatcc
     3601 ccgatttctt taagcagtcc ttccctgagg gcttcacatg ggagagagtc accacatacg
     3661 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     3721 acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa
     3781 cactcggctg ggaggccaac accgagatgc tgtaccccgc tgacggcggc ctggaaggca
     3841 gaaccgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttcaagacca
     3901 catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggacc
     3961 acagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg
     4021 ctgtggccag atactgcgac ctccctagca aactggggca caaacttaat ggcatggacg
     4081 agctgtacaa gggaggttca ggaggcagcg agtgcatctc catccacgtt ggccaggctg
     4141 gtgtccagat tggcaatgcc tgctgggagc tctactgcct ggaacacggc atccagcccg
     4201 atggccagat gccaagtgac aagaccattg ggggaggaga tgactccttc aacaccttct
     4261 tcagtgagac gggcgctggc aagcacgtgc cccgggctgt gtttgtagac ttggaaccca
     4321 cagtcattgg tgagttgacc tcagtaacct gagatcccag gatgctggga caggaggtct
     4381 gtccaggggc ttctcttgtc actcactcac tccctccgtc cttctctccc tcctccagat
     4441 gaagttcgca ctggcaccta ccgccagctc ttccaccctg agcagctcat cacaggcaag
     4501 gaagatgctg ccaataacta tgcccgaggg cactacacca ttggcaagga gatcattgac
     4561 cttgtgttgg accgaattcg caagctggta agcaccacat ataaatatgc atttaatgtg
     4621 gtgtgatagt tccagtgcaa gttgggtgga gtgactgaca tcattcattc tttggcacct
     4681 accaaaatgt ggaataggct gcttgctata ttaattggac ttctaaatca gatagtccct
     4741 aggttatgga cagtttgtgg atatgtctgt tttgccaatt ccttgtgctt acatcagtga
     4801 gatatggttc gtaatctaaa aagttgaaat agaaattcta agataatgtg tcctggcatt
     4861 aaaatattac atttttttat tcccctacag gctgaccagt gcaccggtct tcagggcttc
     4921 ttggttttcc acagctttgg tgggggaact ggttctgggt tcacctccct gctcatggaa
     4981 cgtctctcag ttgattatgg caagaagtcc aagctggagt tctccattta cccagcaccc
     5041 caggtttcca cagctgtagt tgagccctac aactccatcc tcaccaccca caccaccctg
     5101 gagcactcat cccaatggcg cgccgagctt ggcgtaatca tggtcatagc tgtttcctgt
     5161 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5221 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5281 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5341 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5401 cgttcggctg cggcgag
//
