LOCUS       sequence-215528-        5823 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNA sequence source for amplifying an HDR template to tag endogenous
            human RAB11A gene with BFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5823)
  AUTHORS   Roth TL, Puig-Saus C, Yu R, Shifrut E, Carnevale J, Li PJ, Hiatt J, 
            Saco J, Krystofinski P, Li H, Tobin V, Nguyen DN, Lee MR, Putnam AL,
            Ferris AL, Chen JW, Schickel JN, Pellerin L, Carmody D, 
            Alkorta-Aranburu G, Del Gaudio D, Matsumoto H, Morell M, Mao Y, Cho 
            M, Quadros RM, Gurumurthy CB, Smith B, Haugwitz M, Hughes SH, 
            Weissman JS, Schumann K, Esensten JH, May AP, Ashworth A, Kupfer GM,
            Greeley SAW, Bacchetta R, Meffre E, Roncarolo MG, Romberg N, Herold 
            KC, Ribas A, Leonetti MD, Marson A
  TITLE     Reprogramming human T cell function and specificity with non-viral 
            genome targeting.
  JOURNAL   Nature. 2018 Jul 11. pii: 10.1038/s41586-018-0326-5. doi: 
            10.1038/s41586-018-0326-5.
  PUBMED    29995861
REFERENCE   2  (bases 1 to 5823)
  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: "Nature. 
            2018 Jul 11. pii: 10.1038/s41586-018-0326-5. doi: 
            10.1038/s41586-018-0326-5."
COMMENT     Sequence Label: RAB11A-BFP HDRT Source (pTR 145)
FEATURES             Location/Qualifiers
     source          1..5823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..56
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(94..116)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     216..235
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     429..451
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     443..460
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     444..460
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1702..2397
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     primer_bind     complement(3667..3683)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3667..3683)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3680..3702)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3691..3707
                     /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(3715..3721,3722..3739,3740..3745))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3760..3781
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3898..3915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4069..4657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4149..4168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4828..5619,5620..5688))
                     /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     5451..5470
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5689..5793)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
       61 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
      121 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
      181 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
      241 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
      301 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
      361 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
      421 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacccg
      481 gggagttata ggcatacttg cccacaagtc taaatcgaga gtttgcatat aggaggtagg
      541 aggtgtgaca ataattggat aaaggctttt ctggaatgac aaaattctct tcaaatacag
      601 ggaggccctg cgacatgata tctggcaaat cacttgagtc ttagtatcca aacttccgct
      661 aaaatggaaa ttaccatcct ttgcttggca tgttaaatga aatacttatg taaacggact
      721 tagattagag gcctacaaat aggactcaag ttcttgatga atcttgatga ggagggtggg
      781 cgaagtactc taaatgaaaa tgagatagat tgcgagattt ccggagagga gtgagtgaca
      841 agcgcctacc cgtgcatgaa gctggaggta gaaaattcca gcaaaaggaa ttttcaagtt
      901 tcactgaagc ttttgaatca aggcttgaaa gcccagggtt aaaaaaaagg cagtgaagaa
      961 gctcattaag acaacccccc gccaggtagc agatagcacc tgaaatgcag tgggaaattt
     1021 gtgaggtctt cataggcaag gattaaattt tacatctcca tcttaagatg tgtcatagtt
     1081 gttcaattac ttagtgacca aaaaagtgaa ttaaagatgt tgtcaagatt aaaaaaacaa
     1141 aaaccaatgc aggttccacc gagatttgaa ctcggatcgc tggattcaga gtccagagtg
     1201 ctaaccatta caccatggaa ccgccacgca tgtgtagctg ccttcggctg tctaatcctc
     1261 agagaacccc gcccccatcc acaaacccac cactcacagg cggtcccgcc tggttccagc
     1321 gagccgcttc cggcacggta gctcgagaaa tgagcaagcg gccactaaga ctatggtagc
     1381 taggagttcc aggactcagt ttcccctttg agcctccttt agcgactaaa gcttgaagcc
     1441 ccacgcatct cgactctcgc gcacaccgcc cttgttgggc tcaggggcgg ggcgccgccc
     1501 ccggaagtac ttccccttaa aggctggggc ctgccggaaa tggcgcagcg gcagggaggg
     1561 gctcttcacc cagtccggca gttgaagctc ggcgctcggg ttacccctgc agcgacgccc
     1621 cctggtccca cagataccac tgctgctccc gccctttcgc tcctcggccg cgcaatgggc
     1681 ggatcgggtg ggactagtgg cagcgagctg attaaggaga acatgcacat gaagctgtac
     1741 atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc
     1801 tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc
     1861 ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag
     1921 ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc
     1981 acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc
     2041 ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag
     2101 aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg
     2161 gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc
     2221 aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat
     2281 gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac
     2341 gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaatgga
     2401 accggtgctg gaagtggtac acgcgacgac gagtacgact acctctttaa aggtgaggcc
     2461 atgggctctc gcactctaca cagtcctcgt tcggggaccc gggccactcc cggtggaccc
     2521 tcgtgccggc cacccctgca ctgatatagg cctccctcag cccttccttt ttgtgcggtt
     2581 ccgtctccta cccagctcag cctcttctcc cccgctcaga caggggtccc catcacatgc
     2641 cgctctctga gcgacctctc cataggcctt cgctggcctc agagcccctc cctgcgtgtc
     2701 cttcccctgg cggactgcct tctcccacat cgtcgaattc ctttccccgg gttctacggc
     2761 cccgccgctc ctcccaccat ctctcttttc gggtgtagcg ccccctcccc ctcggcgtac
     2821 acccttccca gctcgcgtcc tctcccgaag cccctctgac gggttcttcg cttccctctt
     2881 ggccttgcct tcggtgcaga ctcccattac aggtcttttt cttatctgcc ctctttcccc
     2941 cctcgagcga gcgtctcttc tgtatctccc ctccacttgg acggccgccc ctctaagctt
     3001 ctcccatccc gggtccagga ccccgtaaaa tctgagcggg ccaagggcct ccgcccccac
     3061 cctccgcccc gcggttctcg cggtccttaa ccttctctga ccgctccctc tcccactccc
     3121 catttggcaa ctctctctac cttccctccc caaatcgtgt ttagagctcc cttgtctccg
     3181 ttgcgacctg ggtggtggga gggagggtaa gagggatcgc ccatcaccca ctcgcgcact
     3241 tgtcgcagtg acccggcagg ctgacattac ttcccaccta gccttttctt ttcttgccct
     3301 ccctactcag tggcacccaa cccgggataa ctctttactc ggttcgcgag atgaggttgc
     3361 ggatgggtgg tggctgttat ccttgccaga cctgaaggcc tggttcctga gcaggaaagg
     3421 ttgtgcgctg cacaaatact agcttgcttt atctctatga actggttgct tcctgctgca
     3481 acgctgggag gatccaacag gaagggaaga gccctagtta gccacatacc tttcccaaaa
     3541 ggttgggggt gggaggaagt tgtgtgctct tttggttaca ttaaatattt attataagat
     3601 acggaaacat gtagtgccct ttcctctaga gtcgacctgc aggcatgcaa gcttggcgta
     3661 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     3721 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     3781 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     3841 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     3901 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3961 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4021 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4081 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4141 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4201 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4261 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4321 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4381 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4441 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4501 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4561 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4621 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4681 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4741 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4801 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4861 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     4921 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     4981 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5041 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5101 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5161 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5221 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5281 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5341 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5401 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5461 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5521 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5581 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5641 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5701 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     5761 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5821 cgt
//
