LOCUS Exported 5472 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pc3XB-ZF16 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5472) TITLE Zinc Finger Consortium Modular Assembly Kit v1.0S REFERENCE 2 (bases 1 to 5472) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5472 /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(1062..1079) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1068..1292 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 1338..1766 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1425..1444) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1635..1656 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1775..1795) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 1780..2109 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 1960..2095 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2022..2041 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2180..2974 /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(2234..2253) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2844..2863 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3148..3269 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3185..3204) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3239..3258 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3318..3334) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3318..3334) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3331..3353) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3342..3358 /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(3366..3396) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3411..3432 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3549..3566) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3720..4305) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3800..3819) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4476..5336) /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 5099..5118 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5337..5441) /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 agccgcatat ttgccacatc caaggctgtg 961 ggaaggtgta cggcacaaca agcgccctga cacgtcactt gcgctggcat accggtggat 1021 cctctgcaca gtagtaagct taagtttaaa ccgctgatca gcctcgactg tgccttctag 1081 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac 1141 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca 1201 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag 1261 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg 1321 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 1381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 1441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 1501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga 1561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 1621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt 1681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 1741 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga 1801 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca 1861 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc 1921 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc 1981 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag 2041 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc 2101 ttttgcaaaa agctcccggc cgggagcttg tatatccatt ttcggatctg atcaagagac 2161 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc 2221 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc 2281 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc 2341 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg 2401 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt 2461 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc 2521 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga 2581 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga 2641 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct 2701 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc 2761 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt 2821 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg 2881 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat 2941 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc 3001 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa 3061 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat 3121 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa 3181 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt 3241 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag 3301 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 3361 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 3421 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 3481 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 3541 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 3601 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 3661 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 3721 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 3781 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 3841 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 3901 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 3961 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 4021 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 4081 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 4141 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 4201 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt 4261 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 4321 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 4381 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 4441 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 4501 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 4561 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 4621 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 4681 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 4741 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 4801 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 4861 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 4921 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 4981 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 5041 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 5101 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 5161 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 5221 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 5281 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 5341 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 5401 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 5461 ccacctgacg tc //