LOCUS Exported 7104 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION CRISPR donor plasmid to tag human transcription factor FOXF2 with GFP. ACCESSION . VERSION . KEYWORDS pFOXF2-donor SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7104) TITLE White lab ENCODE plasmids to tag human transcription factors with GFP REFERENCE 2 (bases 1 to 7104) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7104 /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 3873..3893 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 3906..3950 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /label=S-Tag /translation="KETAAAKFERQHMDS" CDS 3990..4013 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 4041..4760 /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(4086..4107) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4347..4366) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" misc_feature 4798..5383 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(4964..4981) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 5191..5210 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 5465..6268 /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(5528..5547) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6138..6157 /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 tagaagaaca 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 tcggtaccca aaactgaatc cggcccgaag cgacgtgcac 2701 tttatggtcc ccacaccact cggttaacta agaaaagacc cgggcgaatg gacctaacgc 2761 agcccggtgc cgagagggcc cggtccagca gcctctgggg cccagtgcgc agggcactgc 2821 gggccgattg cgccccgggg ccaggaggcg ccgagaaagc aaaagcaaaa gccggcggcg 2881 ggtggaggtc aagcggcccg gtgcaggccg gggaccgggg agccgccccg ggagagcagg 2941 gtccaggcct tctctgtccc cctccagcct ggggctacct cagggactcc tcgcgctgcc 3001 agatgcttcg gaccaaaggg catttctctg cgccctgaag tcccggcttc tctagaggaa 3061 gtctaagacg ttcgggaccc acgaagatgc ctgcggaggg gcaggagaag aggcgtgctt 3121 ttctgcccca tcccttcatc ttgcgattgg gttgttttct atcgtcccct ttggcacagg 3181 gagtcctctg cgctcgctgc tgtgaccggc ccagttagtt gggaaacaag gctcagaaag 3241 aaggccgcag tggtgtttct ggggtagggt gcttggccct ccctgaagcc agggctcagg 3301 gcagcacagg ccaggagctc tgccgggaga gacccggaac cagagtctgg gtctagttgt 3361 ccggtacctg tgctcattct gagattaaag tgcacgcccc agcctcagag aataaaattg 3421 gaattccagg acataagaat ccccctcgag ggggaacttg ctctgtgttt gtgcgagcat 3481 caaaacaaaa cataatgatt ccaaagcaat tctatgacgt gcagttggtg tgtttgagct 3541 acttctaaag ctgggctttc tctaccagct gtctagcaat ttactaaaag gcacagcagc 3601 accttttgta ctctgaggca aaataagaca ccctgccatc cactggatga ctttgtttct 3661 gaagaggttt ttttttcttt cttctttcag tgggactgcc ccgttaccag catcactcta 3721 ctccagtgtg tgacagaaaa gatttcgtcc tcaacttcaa tgggatttct tctttccatc 3781 cctcagctag cgggtcgtat tatcaccatc accaccagag cgtctgtcag gatattaagc 3841 cctgcgtcat ggggattcca actactgcaa gcgagaatct ttattttcag ggcgccgcca 3901 aattcaaaga aaccgctgct gctaaattcg aacgccagca catggacagc ggaggtggag 3961 gttcgagcgg tccctcgggt tcgtcgagcc tggaagttct gttccagggg cccctgtcgt 4021 cgagcggtcc ctcgggttcg atggtgagca aaggcgagga gctgttcacc ggggtggtgc 4081 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg 4141 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc 4201 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc 4261 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg 4321 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga 4381 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg 4441 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca 4501 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg 4561 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg 4621 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg 4681 agaagcgcga tcacatggtc cttaaggagt tcgtgaccgc cgccgggatc actctcggca 4741 tggacgagct gtacaagtaa tgatctagag tcgagttaat taagaattcc gccccccccc 4801 ctctccctcc ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg 4861 tttgtctata tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa 4921 cctggccctg tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg 4981 cagggtctgt tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca 5041 acgtctgtag cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc 5101 ggccaaaagc cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt 5161 gtgagttgga tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg 5221 ctgaaggatg cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca 5281 tgctttacat gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac 5341 gtggttttcc tttgaaaaac acgatgataa tatggccaca acctgttaca ttgcacaaga 5401 taaaaatata tcatcacgaa cagtaaaact gtctgcttac ataaacagta atacaagggg 5461 tgttatggga tcggccattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 5521 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 5581 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 5641 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 5701 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 5761 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 5821 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 5881 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 5941 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 6001 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 6061 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 6121 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 6181 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 6241 ctatcgcctt cttgacgagt tcttctgagt cacggaaaga ggccaagcga tggccgctct 6301 ctcctctccc ctcctcagag ggggcagata gaaactggga cggattcaag tcacatgcac 6361 gcggatagca gtaagccaca cacctgccac ttagccagaa tgcccaggat cgcgttggtc 6421 actgttattt gcctactgct ggaagaagga caaccgctgg caaggtagcg ttccccaatc 6481 tgaatacctg caggctccca catgagggag agggcagact caggtgggaa gatgtgccat 6541 gcgtaaggca tcaacgtgta tctgtgggat cttcgttgcc ttcagtaatc agggtgtgaa 6601 aaaagcagac aagttgtgtg tgtgtgtgtg tgtctaagaa aacttgtgtg cttttcaaaa 6661 aggcagtgct aagcacaaga tttcaagaaa gcctcttctt gttgcctagc tgagtgggag 6721 agtcattttc cccagacact acatttggat acaggtgcca aagaacatta ttaaggaatt 6781 atttagaaac aatgtgtcta gtttaagaaa gtggttttca gtattgtgac aatacaacgt 6841 ttttacaagg ttgttttcta ccaccatatt ttaaagatat ttttatgacc gtgtatactc 6901 acactttgct tgtattttaa aaggaggata tatttgcact tatgtatact tttacagttt 6961 gccaaaatat tttgttgtaa aatttttttc aataaaatgt atataacaaa tagttgtttt 7021 aggggctgat tttctcttcg ggcaccctga gagcaatgga tgccagtttc tcctccccgg 7081 caggcagctg tgcaactcta ccca //