LOCUS       sequence-210642-        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     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1637..2332
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     primer_bind     complement(3602..3618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3602..3618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3615..3637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3626..3642
                     /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(3650..3656,3657..3674,3675..3680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3695..3716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3833..3850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4004..4592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4084..4103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4763..5554,5555..5623))
                     /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     5386..5405
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5624..5728)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5796..5814
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggggag
      421 ttataggcat acttgcccac aagtctaaat cgagagtttg catataggag gtaggaggtg
      481 tgacaataat tggataaagg cttttctgga atgacaaaat tctcttcaaa tacagggagg
      541 ccctgcgaca tgatatctgg caaatcactt gagtcttagt atccaaactt ccgctaaaat
      601 ggaaattacc atcctttgct tggcatgtta aatgaaatac ttatgtaaac ggacttagat
      661 tagaggccta caaataggac tcaagttctt gatgaatctt gatgaggagg gtgggcgaag
      721 tactctaaat gaaaatgaga tagattgcga gatttccgga gaggagtgag tgacaagcgc
      781 ctacccgtgc atgaagctgg aggtagaaaa ttccagcaaa aggaattttc aagtttcact
      841 gaagcttttg aatcaaggct tgaaagccca gggttaaaaa aaaggcagtg aagaagctca
      901 ttaagacaac cccccgccag gtagcagata gcacctgaaa tgcagtggga aatttgtgag
      961 gtcttcatag gcaaggatta aattttacat ctccatctta agatgtgtca tagttgttca
     1021 attacttagt gaccaaaaaa gtgaattaaa gatgttgtca agattaaaaa aacaaaaacc
     1081 aatgcaggtt ccaccgagat ttgaactcgg atcgctggat tcagagtcca gagtgctaac
     1141 cattacacca tggaaccgcc acgcatgtgt agctgccttc ggctgtctaa tcctcagaga
     1201 accccgcccc catccacaaa cccaccactc acaggcggtc ccgcctggtt ccagcgagcc
     1261 gcttccggca cggtagctcg agaaatgagc aagcggccac taagactatg gtagctagga
     1321 gttccaggac tcagtttccc ctttgagcct cctttagcga ctaaagcttg aagccccacg
     1381 catctcgact ctcgcgcaca ccgcccttgt tgggctcagg ggcggggcgc cgcccccgga
     1441 agtacttccc cttaaaggct ggggcctgcc ggaaatggcg cagcggcagg gaggggctct
     1501 tcacccagtc cggcagttga agctcggcgc tcgggttacc cctgcagcga cgccccctgg
     1561 tcccacagat accactgctg ctcccgccct ttcgctcctc ggccgcgcaa tgggcggatc
     1621 gggtgggact agtggcagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     1681 gggcaccgtg gacaaccatc acttcaagtg cacatccgag ggcgaaggca agccctacga
     1741 gggcacccag accatgagaa tcaaggtggt cgagggcggc cctctcccct tcgccttcga
     1801 catcctggct actagcttcc tctacggcag caagaccttc atcaaccaca cccagggcat
     1861 ccccgacttc ttcaagcagt ccttccctga gggcttcaca tgggagagag tcaccacata
     1921 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     1981 ctacaacgtc aagatcagag gggtgaactt cacatccaac ggccctgtga tgcagaagaa
     2041 aacactcggc tgggaggcct tcaccgagac gctgtacccc gctgacggcg gcctggaagg
     2101 cagaaacgac atggccctga agctcgtggg cgggagccat ctgatcgcaa acatcaagac
     2161 cacatataga tccaagaaac ccgctaagaa cctcaagatg cctggcgtct actatgtgga
     2221 ctacagactg gaaagaatca aggaggccaa caacgagacc tacgtcgagc agcacgaggt
     2281 ggcagtggcc agatactgcg acctccctag caaactgggg cacaagctta atggaaccgg
     2341 tgctggaagt ggtacacgcg acgacgagta cgactacctc tttaaaggtg aggccatggg
     2401 ctctcgcact ctacacagtc ctcgttcggg gacccgggcc actcccggtg gaccctcgtg
     2461 ccggccaccc ctgcactgat ataggcctcc ctcagccctt cctttttgtg cggttccgtc
     2521 tcctacccag ctcagcctct tctcccccgc tcagacaggg gtccccatca catgccgctc
     2581 tctgagcgac ctctccatag gccttcgctg gcctcagagc ccctccctgc gtgtccttcc
     2641 cctggcggac tgccttctcc cacatcgtcg aattcctttc cccgggttct acggccccgc
     2701 cgctcctccc accatctctc ttttcgggtg tagcgccccc tccccctcgg cgtacaccct
     2761 tcccagctcg cgtcctctcc cgaagcccct ctgacgggtt cttcgcttcc ctcttggcct
     2821 tgccttcggt gcagactccc attacaggtc tttttcttat ctgccctctt tcccccctcg
     2881 agcgagcgtc tcttctgtat ctcccctcca cttggacggc cgcccctcta agcttctccc
     2941 atcccgggtc caggaccccg taaaatctga gcgggccaag ggcctccgcc cccaccctcc
     3001 gccccgcggt tctcgcggtc cttaaccttc tctgaccgct ccctctccca ctccccattt
     3061 ggcaactctc tctaccttcc ctccccaaat cgtgtttaga gctcccttgt ctccgttgcg
     3121 acctgggtgg tgggagggag ggtaagaggg atcgcccatc acccactcgc gcacttgtcg
     3181 cagtgacccg gcaggctgac attacttccc acctagcctt ttcttttctt gccctcccta
     3241 ctcagtggca cccaacccgg gataactctt tactcggttc gcgagatgag gttgcggatg
     3301 ggtggtggct gttatccttg ccagacctga aggcctggtt cctgagcagg aaaggttgtg
     3361 cgctgcacaa atactagctt gctttatctc tatgaactgg ttgcttcctg ctgcaacgct
     3421 gggaggatcc aacaggaagg gaagagccct agttagccac atacctttcc caaaaggttg
     3481 ggggtgggag gaagttgtgt gctcttttgg ttacattaaa tatttattat aagatacgga
     3541 aacatgtagt gccctttcct ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat
     3601 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3661 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3721 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3781 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3841 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3901 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3961 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4021 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4081 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4141 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4201 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4261 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4321 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4381 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4441 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4501 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4561 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4621 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4681 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4741 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4801 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4861 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     4921 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4981 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5041 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5101 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5161 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5221 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5281 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5341 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5401 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5461 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5521 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5581 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5641 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5701 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5761 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     5821 gtc
//
