LOCUS       Exported                5832 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  DNA sequence source for amplifying an HDR template to tag endogenous
            human RAB11A gene with mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    RAB11A-mCherry HDRT Source (pTR 144)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5832)
  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 5832)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5832
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..2341
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1779..1797)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2047..2066
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3611..3627)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3611..3627)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3624..3646)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3635..3651
                     /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(3659..3689)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3704..3725
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3842..3859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4013..4601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4093..4112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4772..5632)
                     /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     5395..5414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5633..5737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5805..5823
                     /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 atcaggcgtc
      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 agtggcgtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     1681 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     1741 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1801 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     1861 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     1921 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     1981 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2041 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     2101 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     2161 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     2221 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     2281 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     2341 gggaaccggt gctggaagtg gtacacgcga cgacgagtac gactacctct ttaaaggtga
     2401 ggccatgggc tctcgcactc tacacagtcc tcgttcgggg acccgggcca ctcccggtgg
     2461 accctcgtgc cggccacccc tgcactgata taggcctccc tcagcccttc ctttttgtgc
     2521 ggttccgtct cctacccagc tcagcctctt ctcccccgct cagacagggg tccccatcac
     2581 atgccgctct ctgagcgacc tctccatagg ccttcgctgg cctcagagcc cctccctgcg
     2641 tgtccttccc ctggcggact gccttctccc acatcgtcga attcctttcc ccgggttcta
     2701 cggccccgcc gctcctccca ccatctctct tttcgggtgt agcgccccct ccccctcggc
     2761 gtacaccctt cccagctcgc gtcctctccc gaagcccctc tgacgggttc ttcgcttccc
     2821 tcttggcctt gccttcggtg cagactccca ttacaggtct ttttcttatc tgccctcttt
     2881 cccccctcga gcgagcgtct cttctgtatc tcccctccac ttggacggcc gcccctctaa
     2941 gcttctccca tcccgggtcc aggaccccgt aaaatctgag cgggccaagg gcctccgccc
     3001 ccaccctccg ccccgcggtt ctcgcggtcc ttaaccttct ctgaccgctc cctctcccac
     3061 tccccatttg gcaactctct ctaccttccc tccccaaatc gtgtttagag ctcccttgtc
     3121 tccgttgcga cctgggtggt gggagggagg gtaagaggga tcgcccatca cccactcgcg
     3181 cacttgtcgc agtgacccgg caggctgaca ttacttccca cctagccttt tcttttcttg
     3241 ccctccctac tcagtggcac ccaacccggg ataactcttt actcggttcg cgagatgagg
     3301 ttgcggatgg gtggtggctg ttatccttgc cagacctgaa ggcctggttc ctgagcagga
     3361 aaggttgtgc gctgcacaaa tactagcttg ctttatctct atgaactggt tgcttcctgc
     3421 tgcaacgctg ggaggatcca acaggaaggg aagagcccta gttagccaca tacctttccc
     3481 aaaaggttgg gggtgggagg aagttgtgtg ctcttttggt tacattaaat atttattata
     3541 agatacggaa acatgtagtg ccctttcctc tagagtcgac ctgcaggcat gcaagcttgg
     3601 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     3661 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     3721 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     3781 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3841 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3901 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3961 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4021 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4081 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4141 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4201 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4261 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4321 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4381 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4441 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4501 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4561 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4621 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4681 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4741 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4801 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4861 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4921 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4981 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5041 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5101 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5161 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5221 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5281 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5341 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5401 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5461 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5521 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5581 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5641 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5701 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5761 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5821 ggccctttcg tc
//
