LOCUS Exported 5466 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pc3XB-ZF73 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5466) TITLE Zinc Finger Consortium Modular Assembly Kit v1.0B REFERENCE 2 (bases 1 to 5466) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5466 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1056..1073) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1062..1286 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1332..1760 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1419..1438) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1629..1650 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1769..1789) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1774..2103 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1954..2089 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2016..2035 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2174..2968 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2228..2247) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2838..2857 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3142..3263 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3179..3198) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3233..3252 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3312..3328) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3312..3328) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3325..3347) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3336..3352 /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(3360..3390) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3405..3426 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3543..3560) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3714..4299) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3794..3813) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4470..5330) /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 5093..5112 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5331..5435) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaacgg gccctctaga cccggggaga agccttacaa atgcccagaa tgtggaaaga 961 gttttagcga ttgtagggat ctcgctagac accagagaac acataccggt ggatcctctg 1021 cacagtagta agcttaagtt taaaccgctg atcagcctcg actgtgcctt ctagttgcca 1081 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 1141 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 1201 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 1261 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 1321 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 1381 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 1441 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 1501 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 1561 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 1621 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 1681 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 1741 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 1801 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 1861 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 1921 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 1981 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 2041 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 2101 aaaaagctcc cggccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg 2161 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt 2221 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 2281 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 2341 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 2401 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 2461 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 2521 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 2581 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 2641 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 2701 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 2761 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 2821 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 2881 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 2941 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa 3001 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 3061 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 3121 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 3181 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 3241 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 3301 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 3361 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 3421 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 3481 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 3541 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 3601 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 3661 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 3721 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 3781 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 3841 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 3901 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 3961 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 4021 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 4081 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 4141 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 4201 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tttttttgtt 4261 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 4321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 4381 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 4441 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 4501 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 4561 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 4621 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 4681 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 4741 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 4801 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 4861 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 4921 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 4981 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 5041 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 5101 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 5161 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 5221 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 5281 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 5341 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 5401 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 5461 gacgtc //