LOCUS Exported 7480 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION KO of PCC 11901 fadD gene/expresses codon-optimized 'tesA gene from E. coli from inducible cLac143 promoter. ACCESSION . VERSION . KEYWORDS pSW068 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7480) TITLE Unprecedented biomass and fatty acid production by the newly discovered cyanobacterium Synechococcus sp. PCC 11901 JOURNAL Unpublished REFERENCE 2 (bases 1 to 7480) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7480 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(1..105) /gene="bla" /label=AmpR promoter primer_bind 173..191 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(229..251) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 351..370 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 564..586 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 578..595 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 579..595 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 1467..1486 /label=lac operator (symmetric) /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). The symmetric lac operator was optimized for tight binding of lac repressor." protein_bind 1571..1587 /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)." primer_bind complement(2256..2274) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2270..2317 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS 2479..3561 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDFSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind complement(2498..2517) /label=LacI-R /note="LacI, reverse primer" primer_bind complement(3729..3748) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 3740..4549 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(3807..3826) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind complement(5459..5475) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5459..5475) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5472..5494) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5483..5499 /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(5507..5537) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5552..5573 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5690..5707) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5861..6449) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5941..5960) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6620..7480) /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 7243..7262 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata 61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa 121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg 181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga 361 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag 421 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc 481 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 541 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagttaatt 601 aaaatttgac tgcgatcgcc tgcaatggcc tgggcctggg cgacaatttt cgggctaggt 661 tcgtaatctt tgggggtagc gatgcgaata ttcatgccga ctaacgcgcc accgaggagg 721 atcgagtggg cgacattatt gccatcaccc acataggtca tggtcaggcc ttcgagttgg 781 ccgaaggttt cctgcatcgt ttgcagatcc gccaacatct gacaggggtg ttctaaatct 841 gtaagggcgt tgataatcgg catttcacag taatcagcga aggtttcgag atcctgttga 901 tcaaaggtac gaatcgccaa aatatcgaga tagcggtcta aaaccctggc ggtgtcctcg 961 atgggctcgc cgcgacccac ttgggtacgg tttgggttca ggtcaatgac attgccaccc 1021 agttggtaca tggccacgct aaaggagact cgggtgcggg tagaggcttt gtaaaataac 1081 agaccgaggg ttttggggca ggtgggggcg atttttccct gtttgagatc tgccgccagg 1141 tttagcaggg taataatttc tgtgctgctg agatctgcaa tgctcagaag atcccgaccc 1201 ttgagtgcat tcatcctgtt ttcggctcct taccgtggca aaaaaatcat tataagctcc 1261 cctccagatt ccacaaaagc ttgcaagggc gatcgccact ggtcttcgtc aggaaatgtt 1321 gatggcgatc ccaagggggt gacacttgaa aaactgtcta cttttgggga gatgggtgaa 1381 atctggtttt aaagtaggat tgtagccatg atttcggctt gttttttgcg cttttaaatg 1441 gaattgcctc tggacatctc caaacgaatt gtgagcgctc acaattcgga attcttaaca 1501 aaaaagcagg aataaaatta acaagatgta attgacataa gtcccatcac cgttgtataa 1561 atgtgtggaa ttgtgagcgg ataacaattt cacacaccaa ctcataaagt caagtaggag 1621 attaattcca tggataccct cctcattctc ggcgatagtc tctccgccgg ttatcgcatg 1681 tctgcctctg ccgcctggcc cgccctcctc aatgataaat ggcaaagcaa aacctctgtg 1741 gtgaatgcca gtatttccgg cgatacctct caacaaggcc tcgcccgtct ccccgccctc 1801 ctcaaacaac accaaccccg ttgggtgttg gtggaactcg gcggtaatga tggcctccgc 1861 ggttttcaac cccaacaaac cgaacaaacc ctccggcaaa ttctccaaga tgtgaaagcc 1921 gccaatgccg aacccctcct catgcaaatt cgcctccccg ccaattatgg tcgccgctac 1981 aatgaagcct ttagtgccat ttatcccaaa ctcgccaaag aatttgatgt gcccctcctc 2041 ccctttttca tggaagaagt gtacctcaaa ccccaatgga tgcaagatga tggtattcac 2101 cccaatcggg atgcccaacc ctttattgcc gattggatgg ccaaacaact ccaacccctc 2161 gtgaatcatg atagttaatc tagatttctt gtacaaactc ggccctatcc ggctgtctaa 2221 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc 2281 ccttggggcc tctaaacggg tcttgacggg ttttttgtct agatcaacgg cctcaaccta 2341 ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga tcccggacac 2401 catcgaatgg cgcatttaca gctagctcag tcctaggtac aatgctagcc ggaggagggt 2461 caattcatgg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt 2521 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg 2581 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa 2641 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac 2701 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg 2761 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt 2821 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt 2881 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca 2941 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg 3001 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg 3061 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg 3121 gaacgggaag gcgactttag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat 3181 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg 3241 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac 3301 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc 3361 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag 3421 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg 3481 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc 3541 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc 3601 accgggatct cgaccgatgc ccttgagaaa atgacgagcg aattagtcat ctttctagat 3661 taacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa 3721 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa 3781 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc 3841 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca 3901 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac 3961 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt 4021 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc 4081 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt 4141 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat 4201 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga 4261 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca 4321 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga 4381 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct 4441 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc 4501 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaag cgatccgaat 4561 ggcggaatct tcgggtattg atgctaccct gaaaatattt tgcagatagg gcccctttag 4621 cccactgatt taacaaaacc gagttacatt tctctaggaa aacattcaca tggacggaaa 4681 ccaagtgcct actgttaccc tcaaacgccc aattaatctg aaggtcatcg tgaccccccg 4741 ctggaaagag gaagtacagc agcaactcca gggacaaata gcgacttttg atggtcaatt 4801 acaacaactg gatgtccaag gaaatcgcac cattagtgaa cttcaacaac agggcaatac 4861 cccccaggtc gctcaacaaa tggaaaatat tcaactccag gttaaccaaa aaaagcggga 4921 actccttgag aagaaaaacc aagtgctcca gcagctccaa caggtacaaa acttagaatt 4981 agatcaagag tttctccagg gtcagatgga gagtttcttt gaagtgaaag tgggcgataa 5041 tctggtgcgc aagctgaatg ttgaacttgt ggtccgggat ggtgttgttg aagaaattcg 5101 cggcgacctc taattttttt gtgattgctg tcgcgtgaca gtgttcggct cccccgatga 5161 cacttgcggt gcgggggatt tttaatgggg aaaaaattca tgaaaattgg cattgtcggg 5221 ctgggtttaa tcggtggatc tttggcgatc gccttccggg aaaaaggctt agaagtgttg 5281 ggtatttccc ggaaaaatac aacctgcgaa acggccttga ggaagggaat tgtcgaccaa 5341 gccagcacag atttttctct tctcaaggat acggatgtgg tgtttctctg cacgccgatc 5401 aaagcgattt tgcccacggc tcaaaccctt attccccacc tcaaatccac ggcgatttgt 5461 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 5521 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 5581 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 5641 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 5701 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 5761 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 5821 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 5881 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 5941 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 6001 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 6061 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 6121 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 6181 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 6241 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 6301 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 6361 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 6421 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 6481 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 6541 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 6601 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 6661 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 6721 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 6781 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 6841 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 6901 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 6961 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 7021 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 7081 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 7141 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 7201 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 7261 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 7321 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 7381 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 7441 aaaagggaat aagggcgaca cggaaatgtt gaatactcat //