LOCUS       sequence-210624-        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     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..2149
                     /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(1676..1697)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2284..2305
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3593..3609)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3593..3609)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3606..3628)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3617..3633
                     /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(3641..3647,3648..3665,3666..3671))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3686..3707
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3824..3841)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3995..4583)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4075..4094)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4754..5545,5546..5614))
                     /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     5377..5396
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5615..5719)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5787..5805
                     /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 agtggcagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1681 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg cgcggcgagg gcgagggcga
     1741 tgccaccaac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1801 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1861 ccacatgaag cgccacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1921 caccatcagc ttcaaggacg acggcaccta caagacccgc gccgaggtga agttcgaggg
     1981 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2041 cctggggcac aagctggagt acaacttcaa cagccacaac gtctatatca ccgccgacaa
     2101 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacgtggagg acggcagcgt
     2161 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2221 cgacaaccac tacctgagca cccagtccgt gctgagcaaa gaccccaacg agaagcgcga
     2281 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actggaaccg gtgctggaag
     2341 tggtacacgc gacgacgagt acgactacct ctttaaaggt gaggccatgg gctctcgcac
     2401 tctacacagt cctcgttcgg ggacccgggc cactcccggt ggaccctcgt gccggccacc
     2461 cctgcactga tataggcctc cctcagccct tcctttttgt gcggttccgt ctcctaccca
     2521 gctcagcctc ttctcccccg ctcagacagg ggtccccatc acatgccgct ctctgagcga
     2581 cctctccata ggccttcgct ggcctcagag cccctccctg cgtgtccttc ccctggcgga
     2641 ctgccttctc ccacatcgtc gaattccttt ccccgggttc tacggccccg ccgctcctcc
     2701 caccatctct cttttcgggt gtagcgcccc ctccccctcg gcgtacaccc ttcccagctc
     2761 gcgtcctctc ccgaagcccc tctgacgggt tcttcgcttc cctcttggcc ttgccttcgg
     2821 tgcagactcc cattacaggt ctttttctta tctgccctct ttcccccctc gagcgagcgt
     2881 ctcttctgta tctcccctcc acttggacgg ccgcccctct aagcttctcc catcccgggt
     2941 ccaggacccc gtaaaatctg agcgggccaa gggcctccgc ccccaccctc cgccccgcgg
     3001 ttctcgcggt ccttaacctt ctctgaccgc tccctctccc actccccatt tggcaactct
     3061 ctctaccttc cctccccaaa tcgtgtttag agctcccttg tctccgttgc gacctgggtg
     3121 gtgggaggga gggtaagagg gatcgcccat cacccactcg cgcacttgtc gcagtgaccc
     3181 ggcaggctga cattacttcc cacctagcct tttcttttct tgccctccct actcagtggc
     3241 acccaacccg ggataactct ttactcggtt cgcgagatga ggttgcggat gggtggtggc
     3301 tgttatcctt gccagacctg aaggcctggt tcctgagcag gaaaggttgt gcgctgcaca
     3361 aatactagct tgctttatct ctatgaactg gttgcttcct gctgcaacgc tgggaggatc
     3421 caacaggaag ggaagagccc tagttagcca catacctttc ccaaaaggtt gggggtggga
     3481 ggaagttgtg tgctcttttg gttacattaa atatttatta taagatacgg aaacatgtag
     3541 tgccctttcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc
     3601 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     3661 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     3721 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     3781 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3841 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3901 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3961 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4021 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4081 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4141 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4201 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4261 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4321 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4381 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     4441 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4501 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4561 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4621 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4681 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4741 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4801 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4861 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4921 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4981 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5041 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5101 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5161 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5221 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5281 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5341 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     5401 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     5461 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5521 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5581 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5641 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5701 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5761 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
