LOCUS Exported 6708 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3 HA human NEMO SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6708) AUTHORS Tang ED, Nunez G, Barr FG, Guan KL TITLE Negative regulation of the forkhead transcription factor FKHR by Akt. JOURNAL J Biol Chem. 1999 Jun 11. 274(24):16741-6. PUBMED 10358014 REFERENCE 2 (bases 1 to 6708) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6708 /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" CDS 903..929 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" promoter complement(2261..2279) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2262..2279) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2299..2316) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2305..2529 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2575..3003 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2662..2681) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2872..2893 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3012..3032) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3017..3346 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3197..3332 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3259..3278 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3413..4207 /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(3467..3486) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4077..4096 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4381..4502 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4418..4437) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4472..4491 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4551..4567) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4551..4567) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4564..4586) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4575..4591 /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(4599..4629) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4644..4665 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4782..4799) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4953..5541) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5033..5052) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5712..6572) /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 6335..6354 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6573..6677) /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 ggagacccag cttaccatgg 901 cctaccccta cgacgtgccc gactacgcct ccctcggatc catgaatagg cacctctgga 961 agagccaact gtgtgagatg gtgcagccca gtggtggccc ggcagcagat caggacgtac 1021 tgggcgaaga gtctcctctg gggaagccag ccatgctgca cctgccttca gaacagggcg 1081 ctcctgagac cctccagcgc tgcctggagg agaatcaaga gctccgagat gccatccggc 1141 agagcaacca gattctgcgg gagcgctgcg aggagcttct gcatttccaa gccagccaga 1201 gggaggagaa ggagttcctc atgtgcaagt tccaggaggc caggaaactg gtggagagac 1261 tcggcctgga gaagctcgat ctgaagaggc agaaggagca ggctctgcgg gaggtggagc 1321 acctgaagag atgccagcag cagatggctg aggacaaggc ctctgtgaaa gcccaggtga 1381 cgtccttgct cggggagctg caggagagcc agagtcgctt ggaggctgcc actaaggaat 1441 gccaggctct ggagggtcgg gcccgggcgg ccagcgagca ggcgcggcag ctggagagtg 1501 agcgcgaggc gctgcagcag cagcacagcg tgcaggtgga ccagctgcgc atgcagggcc 1561 agagcgtgga ggccgcgctc cgcatggagc gccaggccgc ctcggaggag aagaggaagc 1621 tggcccagtt gcaggtggcc tatcaccagc tcttccaaga atacgacaac cacatcaaga 1681 gcagcgtggt gggcagtgag cggaagcgag gaatgcagct ggaagatctc aaacagcagc 1741 tccagcaggc cgaggaggcc ctggtggcca aacaggaggt gatcgataag ctgaaggagg 1801 aggccgagca gcacaagatt gtgatggaga ccgttccggt gctgaaggcc caggcggata 1861 tctacaaggc ggacttccag gctgagaggc aggcccggga gaagctggcc gagaagaagg 1921 agctcctgca ggagcagctg gagcagctgc agagggagta cagcaaactg aaggccagct 1981 gtcaggagtc ggccaggatc gaggacatga ggaagcggca tgtcgaggtc tcccaggccc 2041 ccttgccccc cgcccctgcc tacctctcct ctcccctggc cctgcccagc cagaggagga 2101 gcccccccga ggagccacct gacttctgct gtcccaagtg ccagtatcag gcccctgata 2161 tggacaccct gcagatacat gtcatggagt gcattgagta gaattctgca gatatccatc 2221 acactggcgg ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg 2281 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2341 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2401 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2461 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 2521 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 2581 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 2641 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 2701 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 2761 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 2821 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 2881 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 2941 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 3001 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3061 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3181 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3241 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 3301 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 3361 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 3421 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 3481 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 3541 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 3601 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 3661 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 3721 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 3781 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 3841 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 3901 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 3961 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 4021 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 4081 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 4141 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4201 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 4261 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4321 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 4381 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4441 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4501 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 4561 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 4621 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 4681 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 4741 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 4801 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4861 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 4921 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 4981 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 5041 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5101 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 5161 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5221 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5281 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 5341 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 5401 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 5461 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 5521 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 5581 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 5641 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 5701 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 5761 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 5821 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 5881 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 5941 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 6001 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 6061 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 6121 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 6181 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 6241 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 6301 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 6361 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 6421 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 6481 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 6541 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 6601 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 6661 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc //