LOCUS Exported 7449 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS PKCalpha.FLAG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7449) AUTHORS Hodges RR, Kazlauskas A, Toker A, Dartt DA TITLE Effect of overexpression of protein kinase C alpha on rat lacrimal gland protein secretion. JOURNAL Adv Exp Med Biol. 2002 . 506(Pt A):237-41. PUBMED 12613914 REFERENCE 2 (bases 1 to 7449) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7449 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 171..550 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 551..754 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 705..725 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 799..818 /label=T7 /note="T7 promoter, forward primer" promoter 799..817 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 2896..2919 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" promoter complement(2938..2956) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2939..2956) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2976..2993) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2982..3206 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3252..3680 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3339..3358) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3549..3570 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3689..3709) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3694..4023 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3874..4009 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3936..3955 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4090..4884 /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(4144..4163) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4754..4773 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5058..5179 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5095..5114) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5149..5168 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5228..5244) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5228..5244) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5241..5263) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5252..5268 /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(5276..5306) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5321..5342 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5459..5476) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5630..6218) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5710..5729) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6389..7249) /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 7012..7031 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7250..7354) /gene="bla" /label=AmpR promoter primer_bind complement(7429..7448) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag 61 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 121 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt 181 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 241 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 301 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 361 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 421 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 481 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 541 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 601 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 661 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 721 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc 781 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc 841 tcggatccac tagtaacggc cgccagtgtg ctggaattca tggctgacgt cttcccggcc 901 gccgagccgg cggcgccgca ggacgtggcc aaccgcttcg cccgcaaagg ggcgctgagg 961 cagaagaacg tgcacgaggt gaagaaccac cgcttcatcg cgcgcttctt caagcagccc 1021 accttctgca gccactgcac cgacttcatc tgggggtttg ggaaacaagg cttccagtgc 1081 caagtttgct gttttgtggt tcacaagagg tgccatgaat ttgttacttt ttcttgtccg 1141 ggggcggata aaggacccga cacagatgac ccgaggagca agcacaagtt caagatccac 1201 acgtatggca gccccacctt ctgtgatcac tgcggctccc tgctgtacgg actcatccac 1261 caggggatga aatgtgacac ctgtgatatg aacgtgcaca agcagtgcgt gatcaacgtg 1321 cccagcctct gcgggatgga ccacacggag aagaggggcc gcatctacct gaaggccgag 1381 gtcacggatg aaaagttgca cgtcacagta cgagacgcga aaaacctaat ccctatggat 1441 ccaaatgggc tttcagatcc ttacgtgaag ctgaagctta ttcctgaccc caagaacgag 1501 agcaaacaga aaaccaagac catccgctcg acgctgaacc cccggtggga cgagtccttc 1561 acgttcaaat taaaaccttc tgataaagac cggcgactgt ccgtggaaat ctgggactgg 1621 gatcgaacca cacggaacga cttcatgggg tccctttcct ttggggtctc ggagctgatg 1681 aagatgccgg ccagcggatg gtacaagctg ctgaaccaag aggagggcga gtactacaac 1741 gtgccgatcc ccgaaggcga cgaggaaggc aatgtggagc tcaggcagaa attcgagaaa 1801 gccaagcttg gccctgccgg caacaaagtc atcagtccct ccgaggacag gagacagcct 1861 tccaacaacc tgggcagagt gaagctcacg gacttcaact tcctcatggt gctgggcaaa 1921 ggcagctttg ggaaggtgat gctggccgac cgaaagggga cagaggagct gtacgccatc 1981 aagatcctga agaaggacgt ggtcatccag gacgacgacg tggagtgcac catggtggag 2041 aagcgggtcc tggcgctgct cgacaagccg ccgttcctga cgcagctgca ctcctgcttc 2101 cagacggtgg accggctgta cttcgtcatg gagtacgtca acggcgggga cctcatgtac 2161 cacatccagc aggtcgggaa gttcaaggag ccgcaagcag tgttctatgc agcagagatt 2221 tccatcgggc tgttctttct tcataaaaga ggaatcattt atcgggacct gaagttagac 2281 aacgtcatgc tggactcgga aggacacatt aagatcgcgg acttcgggat gtgcaaggag 2341 cacatgatgg acggcgtcac gaccaggacc ttctgcggga cccccgacta catcgcccca 2401 gagataatcg cctatcagcc gtacgggaag tccgtggact ggtgggccta cggcgtcctg 2461 ttgtacgaga tgttggccgg gcagcctccg ttcgacggcg aggacgagga cgagctgttc 2521 cagtccatca tggagcacaa cgtctcgtac cccaagtcct tgtccaagga ggccgtgtcc 2581 atctgcaaag ggctgatgac caagcacccc gggaagcggc tgggctgcgg gcccgagggc 2641 gagcgcgacg tgcgggagca tgccttcttc cggaggatcg actgggagaa gctggagaac 2701 cgtgagatcc agccaccctt caagcccaaa gtgtgcggca aaggagcaga gaactttgac 2761 aagttcttca cgcgagggca gcctgtcttg acgccgcccg accagctggt catcgctaac 2821 atcgaccagt ctgattttga aggcttctcc tacgtcaacc cccagttcgt gcaccccatc 2881 ctgcagagcg cggtagacta taaggacgat gatgacaaat gatctagagg gccctattct 2941 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca 3001 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac 3061 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat 3121 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca 3181 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag 3241 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 3301 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 3361 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 3421 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 3481 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 3541 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 3601 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 3661 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc 3721 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 3781 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 3841 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 3901 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 3961 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 4021 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga 4081 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 4141 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 4201 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 4261 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 4321 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 4381 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 4441 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 4501 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 4561 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 4621 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 4681 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 4741 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 4801 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 4861 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 4921 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct 4981 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg 5041 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata 5101 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 5161 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt 5221 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca 5281 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat 5341 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 5401 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 5461 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 5521 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 5581 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 5641 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 5701 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 5761 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 5821 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 5881 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 5941 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 6001 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 6061 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 6121 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 6181 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 6241 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 6301 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 6361 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 6421 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 6481 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct 6541 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 6601 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 6661 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 6721 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 6781 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 6841 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 6901 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 6961 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 7021 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 7081 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 7141 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 7201 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 7261 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 7321 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg 7381 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc 7441 tgatgccgc //