LOCUS Exported 7438 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS eIF2a 2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7438) TITLE eIF2-alpha constructs REFERENCE 2 (bases 1 to 7438) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7438 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(42..61) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 233..612 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 613..816 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 767..787 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 861..880 /label=T7 /note="T7 promoter, forward primer" promoter 861..879 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(2603..2621) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(2604..2621) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2641..2658) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2647..2871 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2917..3345 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3004..3023) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3214..3235 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3354..3374) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3359..3688 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3539..3674 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3601..3620 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5109..5230 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5146..5165) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5200..5219 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5279..5295) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5279..5295) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5292..5314) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5303..5319 /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(5327..5357) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5372..5393 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5510..5527) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5681..6269) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5761..5780) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6440..7300) /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 7063..7082 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7301..7405) /gene="bla" /label=AmpR promoter ORIGIN 1 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc 61 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg 121 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt 181 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg ttgacattga 241 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg 301 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc 361 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat 421 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat 481 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat 541 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc 601 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac 661 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa 721 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt 781 aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag agaacccact 841 gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc ttggtaccga 901 gctcggatcc actagtaacg gccgccagtg tgctggaatt ccggggaagc aagtctggtc 961 tctgtgattg aagaagtcgg ctctgggctc cagtgcggga atcacacaca tacctcagaa 1021 tgccgggtct aagttgtaga ttttatcaac acaaatttcc tgaggtggaa gatgtagtga 1081 tggtgaatgt cagatccatt gctgaaatgg gggcttatgt cagcttgctg gaatacaaca 1141 acattgaagg catgattctt cttagtgaat tggccagaag gcgtatccgt tctatcaaca 1201 aactcatccg aattggcagg aatgagtgtg tggttgtcat tagggtggac aaagaaaaag 1261 gatatattga tttgtcaaaa agaagagttt ctccagagga agcaatcaaa tgtgaagaca 1321 aattcacaaa atccaaaact gtttatagca ttcttcgtca tgttgctgag gtgttagaat 1381 acaccaagga tgagcagctg gaaagcctat tccagaggac tgcctgggtc tttgatgaca 1441 agtacaagag acctggatat ggtgcctatg atgcatttaa gcatgcagtc tcagacccat 1501 ctattttgga tagtttagat ttgaatgaag atgaacggga agtactcatt aataatatta 1561 ataggcgctt gaccccacag gctgtcaaaa ttcgagcaga tattgaagtg gcttgttatg 1621 gttatgaagg cattgatgct gtaaaagaag ccctaagagc aggtttgaat tgttctacag 1681 aaaacatgcc cattaagatt aatctaatag ctcctcctcg gtatgtaatg actacgacaa 1741 ccctggagag aacagaaggc ctttctgtcc tcagtcaagc tatggctgtt atcaaagaga 1801 agattgagga aaagaggggt gtgttcaatg ttcaaatgga gcccaaagtg gtcacagata 1861 cagatgagac tgaacttgcg aggcagatgg agaggcttga aagagaaaat gccgaagtgg 1921 atggagatga tgatgcagaa gaaatggaag ccaaagctga agattaactt tgtgggaaac 1981 agagtccaat ttaaggaaca cagagcagcg cttcctggct gtaaatccta gacttgaaag 2041 ttttccagta ttgaaaactt caaagctgaa tattttttat ttctaagtat ttaaatgttc 2101 taacagatca gaacatgaaa tgccctccta aatgtcagct gttgtcacac agtagctcca 2161 acactttgag catttttaag ggagtggcct catttcacta gagacaaatc tttaagaata 2221 gttctaaaat tgggcttgtg atttccattt ctgatgtctc cagattggca cccctttcta 2281 gttcaatgcc tcacgagatt tgccaggggc atccaaggca aacaatccca atctttctat 2341 ataaaatgta ttcaagcaaa catcaaataa atttctggga tatttaacta taggcttctt 2401 ccttcttgtc accagttaaa agcattttaa tactaagacc ctaattcttt tatctttatt 2461 ttagtcttga tgtggaactg taggagcagg tgaataaagg atctctataa cagatccttt 2521 caaaagaaga gttttagaga aaataaattt aactttaacc acagtgaaag ttgacctcga 2581 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag 2641 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct 2701 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc 2761 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg 2821 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg 2881 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa 2941 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 3001 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 3061 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 3121 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 3181 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 3241 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct 3301 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt 3361 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 3421 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag 3481 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat 3541 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt 3601 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg 3661 cttttttgga ggcctaggct tttgcaaaaa gctcccgggc tcgactagag atccctcgac 3721 ctcgagatcc attgtgctct aaagcctaag atgagctttc catgtaaatt tgtagccagc 3781 ttccttctga ttttcaatgt ttcttccaaa ggtgcagtct ccaaagagat tacgaatgcc 3841 ttggaaacct ggggtgcctt gggtcaggac atcaacttgg acattcctag ttttcaaatg 3901 agtgatgata ttgacgatat aaaatgggaa aaaacttcag acaagaaaaa gattgcacaa 3961 ttcagaaaag agaaagagac tttcaaggaa aaagatacat ataagctatt taaaaatgga 4021 actctgaaaa ttaagcatct gaagaccgat gatcaggata tctacaaggt atcaatatat 4081 gatacaaaag gaaaaaatgt gttggaaaaa atatttgatt tgaagattca agagagggtc 4141 tcaaaaccaa agatctcctg gacttgtatc aacacaaccc tgacctgtga ggtaatgaat 4201 ggaactgacc ccgaattaaa cctgtatcaa gatgggaaac atctaaaact ttctcagagg 4261 gtcatcacac acaagtggac caccagcctg agtgcaaaat tcaagtgcac agcagggaac 4321 aaagtcagca aggaatccag tgtcgagcct gtcagctgtc cagagaaagg tctggacatc 4381 tatctcatca ttggcatatg tggaggaggc agcctcttga tggtctttgt ggcactgctc 4441 gttttctata tcaccaaaag gaaaaaacag aggagtcgga gaaatgatga ggagctggag 4501 acaagagccc acagagtagc tactgaagaa aggggccgga agccccaaca aattccagct 4561 tcaacccctc agaatccagc aacttcccaa catcctcctc caccacctgg tcatcgttcc 4621 caggcaccta gtcatcgtcc cccgcctcct ggacaccgtg ttcagcacca gcctcagaag 4681 aggcctcctg ctccgtcggg cacacaagtt caccagcaga aaggcccgcc cctccccaga 4741 cctcgagttc agccaaaacc tccccatggg gcagcagaaa actcattgtc cccttcctct 4801 aattaaaaaa gatagaaact gtctttttca ataaaaagca ctgtggattt ctgccctcct 4861 gatgtgcata tccgtacttc catgaggtgt tttctgtgtg cagaacattg tcacctcctg 4921 aggctgtggg ccacagccac ctctgcatct tcgaaatgac cgaccaagcg acgcccaacc 4981 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg 5041 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg 5101 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5161 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5221 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt 5281 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5341 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 5401 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 5461 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5521 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5581 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5641 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5701 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5761 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 5821 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 5881 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 5941 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6001 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6061 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6121 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6181 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6241 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6301 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6361 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6421 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6481 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6541 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6601 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6661 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6721 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6781 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 6841 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 6901 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 6961 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7021 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7081 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7141 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7201 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7261 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 7321 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 7381 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcga //