LOCUS       Exported                7206 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  CRISPR donor plasmid to tag human transcription factor HOXC13 with 
            GFP.
ACCESSION   .
VERSION     .
KEYWORDS    pHOXC13-donor
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7206)
  TITLE     White lab ENCODE plasmids to tag human transcription factors with 
            GFP
REFERENCE   2  (bases 1 to 7206)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7206
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(34..50)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(34..50)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(47..69)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    58..74
                     /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(82..112)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    127..148
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(265..282)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(436..1024)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(516..535)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1195..2055)
                     /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     1818..1837
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2056..2160)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2228..2246
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2284..2306)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2406..2425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2619..2641
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2633..2650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2634..2650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3884..3904
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3917..3961
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             4001..4024
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             4052..4771
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4097..4118)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4358..4377)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    4809..5394
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4975..4992)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5202..5221
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5476..6279
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5539..5558)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6149..6168
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
       61 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
      121 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
      181 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
      241 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
      301 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      361 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      421 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      481 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      541 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      601 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      661 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      721 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      781 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      841 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      901 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      961 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1021 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     1081 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     1141 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     1201 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1261 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1321 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1381 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1441 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1501 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1561 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1621 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1681 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1741 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1801 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1861 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1921 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1981 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     2041 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2101 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2161 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2221 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2281 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2401 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     2461 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     2521 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     2581 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     2641 gacggccagt gaattcgagc tcggtacccg acaggcatct agggtttagg gttatggttg
     2701 gtgcaaggga tgtagacagt ttccttcagc tgtgaaggaa tggagaggga aggcggttgg
     2761 aacagaattt gttctgtttt ggtgggactg cagtgtggac cccctacttc cagagacacc
     2821 tgattcccta tatctcaggt tcaggacccc ccaccttggg tgttaggtgg ttcataacat
     2881 tggaccccaa ttcctagagt gtggacctcc aagcaggctg gcttcacctg gacacttatc
     2941 ttctatctgt gcctcacatc cctaacgtag ggccagtgtc caaaggcgtg aagactcatc
     3001 tggtggtctg ggcccactga agctgacccc ctgcctgcca tgccctccct gaaatataca
     3061 acagaatgtc ttcctctggc aacggggcag aatataaact aataaggacc caagcgatct
     3121 aactgcagga aggagggatt tttctgaata tctgaaattc tgcaagcatg gcaggccagg
     3181 ctggaatttg gagggtccct tctaaatata caaggatcct aatattgagt gctgtgaaga
     3241 gccatattgg ggcctgaaac aaaagaaaat cagtaatacc gaacccatgt ttattttttt
     3301 aattgatatt ttgttcatca tggatgtttt tgtattcgtt ttgctttttt aaacattgca
     3361 ttaaaatatc gtcttgatta ctgagttttt ggcgccccct cccttaaact tcgagcccag
     3421 tggcctcatc atagttgtgg tcttgctaaa ataacagaaa tgaaaagcat tagggagctt
     3481 cactatgcag tgcaatcctc tggccccgtt gagcccctgc cagaactctg tgcctaggca
     3541 gcctcgggtc ctctatctca gtccagccgc ttgcctcact tcttcccgct tgccttatct
     3601 ccccgcagac gtggttcccc tgcagcccga ggtgagcagc taccggcgcg ggcgcaagaa
     3661 acgcgtgccc tacactaagg tgcagctgaa ggagctagag aaggaatacg cggctagcaa
     3721 gttcatcacc aaagagaagc gccggcgcat ctccgccacc acgaacctct ctgagcgcca
     3781 ggtaaccatc tggttccaga accggcgggt caaagagaag aaggtggtca gcaaatcgaa
     3841 agcgcctcat ctccactcca ccgggattcc aactactgca agcgagaatc tttattttca
     3901 gggcgccgcc aaattcaaag aaaccgctgc tgctaaattc gaacgccagc acatggacag
     3961 cggaggtgga ggttcgagcg gtccctcggg ttcgtcgagc ctggaagttc tgttccaggg
     4021 gcccctgtcg tcgagcggtc cctcgggttc gatggtgagc aaaggcgagg agctgttcac
     4081 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
     4141 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
     4201 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
     4261 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
     4321 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
     4381 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
     4441 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
     4501 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
     4561 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     4621 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
     4681 agaccccaac gagaagcgcg atcacatggt ccttaaggag ttcgtgaccg ccgccgggat
     4741 cactctcggc atggacgagc tgtacaagta atgatctaga gtcgagttaa ttaagaattc
     4801 cgcccccccc cctctccctc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4861 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4921 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4981 ccaaaggaat gcagggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5041 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5101 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5161 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5221 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5281 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5341 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aacctgttac
     5401 attgcacaag ataaaaatat atcatcacga acagtaaaac tgtctgctta cataaacagt
     5461 aatacaaggg gtgttatggg atcggccatt gaacaagatg gattgcacgc aggttctccg
     5521 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5581 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5641 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5701 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5761 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5821 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5881 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5941 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     6001 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     6061 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     6121 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     6181 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     6241 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag tcccacccac ccgctgcttg
     6301 ccccatctat ttatgtctcc gctttgtacc ataaccgaac ccacggaaag acgctgcgcg
     6361 ggtgcagaag agtatttaat gttaaggaaa gagaagaacc gcgccgcccg gaggcagaga
     6421 ggctccatgg ccgtgctgct gggccatccc caactcccta tcccatcccc agcctccacc
     6481 cccatccaga tgggactcac gtggcttcaa cagctttgga aatgggtccc gagtgggccg
     6541 tgcgaggaag gctgtcgacc tctactcctc cttgcgctca ccttgccaga aagtctggtg
     6601 gcagcgcaga gcccaatcat tccaaccaaa gcagggttgg ggaatcccga atggccccaa
     6661 ttcttgcctc atcctatgac caggctttta gaggaccttc cctaagggcg cagcttcgga
     6721 gcaggatctg tccagctcat actttccttc gctgtccctc ccgcactcct taggcaagat
     6781 ttcccagtaa agattttctg tgcgtatttt aaaagtcgtg ttaatactca tgataattat
     6841 tagggacctg gcagcgtgat tggagtatgg atgtttccgt aaaagctgga attccgtaaa
     6901 agcattgacg cagcccctac actccatccc aaccaagaaa ctgcatttcc tggggccagg
     6961 tgggagctgc ctttgcccca ctgcctcccc tgttctgctc tctcagtcaa catgtggaaa
     7021 tccaaggagg acaaagactc cagccacgct gctaaatagg gctcctctct cctctctctc
     7081 tctctaggtg gtaaggttgg ggattagtcc aggtacagaa gcagaacttt tttctaagga
     7141 taaacatctc ttccaagggg atggagagtg ggtccctcaa caaagtccct gtccagtcac
     7201 ctttcc
//
