LOCUS       sequence-215519-        5814 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DNA sequence source for amplifying an HDR template to tag endogenous
            human RAB11A gene with GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5814)
  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 5814)
  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-GFP HDRT Source (pTR 143)
FEATURES             Location/Qualifiers
     source          1..5814
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     235..254
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(473..577)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     645..663
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(701..723)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     823..842
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1036..1058
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1050..1067
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1051..1067
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2309..2821
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVE"
     primer_bind     complement(2348..2369)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2956..2977
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4265..4281)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4265..4281)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4278..4300)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4289..4305
                     /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(4313..4319,4320..4337,4338..4343))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4358..4379
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4496..4513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4667..5255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4747..4766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5426..5814,1..403,404..472))
                     /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"
ORIGIN
        1 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
       61 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      121 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      181 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      241 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      301 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      361 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      421 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      481 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      541 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      601 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
      661 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
      721 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
      781 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      841 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
      901 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
      961 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     1021 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg
     1081 gtacccgggg agttataggc atacttgccc acaagtctaa atcgagagtt tgcatatagg
     1141 aggtaggagg tgtgacaata attggataaa ggcttttctg gaatgacaaa attctcttca
     1201 aatacaggga ggccctgcga catgatatct ggcaaatcac ttgagtctta gtatccaaac
     1261 ttccgctaaa atggaaatta ccatcctttg cttggcatgt taaatgaaat acttatgtaa
     1321 acggacttag attagaggcc tacaaatagg actcaagttc ttgatgaatc ttgatgagga
     1381 gggtgggcga agtactctaa atgaaaatga gatagattgc gagatttccg gagaggagtg
     1441 agtgacaagc gcctacccgt gcatgaagct ggaggtagaa aattccagca aaaggaattt
     1501 tcaagtttca ctgaagcttt tgaatcaagg cttgaaagcc cagggttaaa aaaaaggcag
     1561 tgaagaagct cattaagaca accccccgcc aggtagcaga tagcacctga aatgcagtgg
     1621 gaaatttgtg aggtcttcat aggcaaggat taaattttac atctccatct taagatgtgt
     1681 catagttgtt caattactta gtgaccaaaa aagtgaatta aagatgttgt caagattaaa
     1741 aaaacaaaaa ccaatgcagg ttccaccgag atttgaactc ggatcgctgg attcagagtc
     1801 cagagtgcta accattacac catggaaccg ccacgcatgt gtagctgcct tcggctgtct
     1861 aatcctcaga gaaccccgcc cccatccaca aacccaccac tcacaggcgg tcccgcctgg
     1921 ttccagcgag ccgcttccgg cacggtagct cgagaaatga gcaagcggcc actaagacta
     1981 tggtagctag gagttccagg actcagtttc ccctttgagc ctcctttagc gactaaagct
     2041 tgaagcccca cgcatctcga ctctcgcgca caccgccctt gttgggctca ggggcggggc
     2101 gccgcccccg gaagtacttc cccttaaagg ctggggcctg ccggaaatgg cgcagcggca
     2161 gggaggggct cttcacccag tccggcagtt gaagctcggc gctcgggtta cccctgcagc
     2221 gacgccccct ggtcccacag ataccactgc tgctcccgcc ctttcgctcc tcggccgcgc
     2281 aatgggcgga tcgggtggga ctagtggcag caagggcgag gagctgttca ccggggtggt
     2341 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgcgcggcga
     2401 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2461 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2521 ccgctacccc gaccacatga agcgccacga cttcttcaag tccgccatgc ccgaaggcta
     2581 cgtccaggag cgcaccatca gcttcaagga cgacggcacc tacaagaccc gcgccgaggt
     2641 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2701 ggacggcaac atcctggggc acaagctgga gtacaacttc aacagccaca acgtctatat
     2761 caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacgtgga
     2821 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2881 cgtgctgctg cccgacaacc actacctgag cacccagtcc gtgctgagca aagaccccaa
     2941 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactggaac
     3001 cggtgctgga agtggtacac gcgacgacga gtacgactac ctctttaaag gtgaggccat
     3061 gggctctcgc actctacaca gtcctcgttc ggggacccgg gccactcccg gtggaccctc
     3121 gtgccggcca cccctgcact gatataggcc tccctcagcc cttccttttt gtgcggttcc
     3181 gtctcctacc cagctcagcc tcttctcccc cgctcagaca ggggtcccca tcacatgccg
     3241 ctctctgagc gacctctcca taggccttcg ctggcctcag agcccctccc tgcgtgtcct
     3301 tcccctggcg gactgccttc tcccacatcg tcgaattcct ttccccgggt tctacggccc
     3361 cgccgctcct cccaccatct ctcttttcgg gtgtagcgcc ccctccccct cggcgtacac
     3421 ccttcccagc tcgcgtcctc tcccgaagcc cctctgacgg gttcttcgct tccctcttgg
     3481 ccttgccttc ggtgcagact cccattacag gtctttttct tatctgccct ctttcccccc
     3541 tcgagcgagc gtctcttctg tatctcccct ccacttggac ggccgcccct ctaagcttct
     3601 cccatcccgg gtccaggacc ccgtaaaatc tgagcgggcc aagggcctcc gcccccaccc
     3661 tccgccccgc ggttctcgcg gtccttaacc ttctctgacc gctccctctc ccactcccca
     3721 tttggcaact ctctctacct tccctcccca aatcgtgttt agagctccct tgtctccgtt
     3781 gcgacctggg tggtgggagg gagggtaaga gggatcgccc atcacccact cgcgcacttg
     3841 tcgcagtgac ccggcaggct gacattactt cccacctagc cttttctttt cttgccctcc
     3901 ctactcagtg gcacccaacc cgggataact ctttactcgg ttcgcgagat gaggttgcgg
     3961 atgggtggtg gctgttatcc ttgccagacc tgaaggcctg gttcctgagc aggaaaggtt
     4021 gtgcgctgca caaatactag cttgctttat ctctatgaac tggttgcttc ctgctgcaac
     4081 gctgggagga tccaacagga agggaagagc cctagttagc cacatacctt tcccaaaagg
     4141 ttgggggtgg gaggaagttg tgtgctcttt tggttacatt aaatatttat tataagatac
     4201 ggaaacatgt agtgcccttt cctctagagt cgacctgcag gcatgcaagc ttggcgtaat
     4261 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgag
//
