LOCUS Exported 3503 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGP-FF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3503) TITLE Zinc Finger Consortium Expression Vector Kit v1.0 REFERENCE 2 (bases 1 to 3503) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3503 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 12..42 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 50..66 /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)." primer_bind 55..77 /label=M13/pUC Reverse /note="In lacZ gene" CDS 368..391 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" rep_origin 564..1019 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(651..670) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 861..882 /label=F1ori-F /note="F1 origin, forward primer" promoter 1045..1149 /gene="bla" /label=AmpR promoter CDS 1150..2010 /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 complement(1368..1387) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2181..2769 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2670..2689 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2923..2940 /label=L4440 /note="L4440 vector, forward primer" misc_feature 2955..3095 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(3021..3040) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3181..3203 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(3197..3388) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" ORIGIN 1 gaattccggg ctttacactt tatgcttccg gctcgtataa tgtgtcgact tgtgagcgga 61 taacaatttc acacaggaaa cagctatgcc tcaacagcag caaatgcaac ctcccaattc 121 aagtgcggac aacaaccctt tgcaacagca atcatcacaa aataccgtac caaacgtcct 181 caaccaaatt aaccaaatct tttctccaga ggagcaacgc agcttattac aagaagccat 241 cgaaacctgc aagaattttg aaaaaacaca attgggtagt acgatgacgg aacctgtcaa 301 gcaaagtttt attaggaaat acattgtgca aaaggccctg agaaaaatcc aagctttggc 361 ggccgctgac tacaaagacg atgatgacaa atctagagtg cagtgataac ctctactcac 421 ctgcgtggat cctctgcaca gtagtaacta gagactagaa aaaggccgac aagtcccgct 481 ccgctgaaga tcctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt 541 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 601 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 661 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 721 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 781 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 841 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 901 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 961 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta 1021 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 1081 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 1141 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt 1201 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag 1261 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt 1321 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg 1381 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag 1441 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta 1501 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg 1561 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta 1621 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac 1681 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt 1741 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca 1801 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag 1861 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta 1921 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag 1981 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt 2041 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 2101 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 2161 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 2221 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 2281 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag 2341 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 2401 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 2461 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 2521 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 2581 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 2641 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 2701 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 2761 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 2821 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 2881 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 2941 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac 3001 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca 3061 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg 3121 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct 3181 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc 3241 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt 3301 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt 3361 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca 3421 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg 3481 aacatgcccg gttactggaa cgg //