LOCUS Exported 6735 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION KO of PCC 7002 fadD gene/expresses codon-optimized 'tesA gene from E. coli from inducible cLac143 promoter. ACCESSION . VERSION . KEYWORDS pSZT025 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6735) TITLE Unprecedented biomass and fatty acid production by the newly discovered cyanobacterium Synechococcus sp. PCC 11901 JOURNAL Unpublished REFERENCE 2 (bases 1 to 6735) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6735 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 38..57 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 251..273 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 265..282 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 266..282 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 779..798 /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 883..899 /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(1568..1586) /label=T7 Term /note="T7 terminator, reverse primer" terminator 1582..1629 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS 1791..2873 /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(1810..1829) /label=LacI-R /note="LacI, reverse primer" primer_bind complement(3022..3041) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 3033..3842 /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(3100..3119) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind complement(4401..4417) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4401..4417) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4414..4436) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4425..4441 /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(4449..4479) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4494..4515 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4632..4649) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4803..5391) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4883..4902) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5562..6422) /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 6185..6204 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6423..6527) /gene="bla" /label=AmpR promoter primer_bind 6595..6613 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6651..6673) /label=pGEX 3' /note="pGEX vectors, reverse primer" ORIGIN 1 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg 61 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc 121 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 181 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 241 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcga gctcggtacc 301 cggggatcct cttcaggtca atgacattgc cacccagttg atacatagcc acgctaaagg 361 agactctggt gcgggtagag gctttgtaga acaacaggcc gagggttttg gggcaggtgg 421 gggcgatttt tccctgtttg agatctgccg ccaggtttag cagggtaata atttctgtgc 481 tgctgagatc tgcaatgctc agaagatccc gacccttgag tgcattcatc ctgttttcgg 541 ctccttaccg tggcaaaaaa atcattataa gctcccctcc agattccaca aaagcttgca 601 agggcgatcg ccactggtct tcgtcaggaa atgttgatgg cgatcccaag ggggtgacac 661 ttgaaaaact gtctactttt ggggagatgg gtgaaatctg gttttaaagt aggattgtag 721 ccatgatttc ggcttgtttt ttgcgctttt aaatggaatt tctggacatc tccaaacgaa 781 ttgtgagcgc tcacaattcg gaattcttaa caaaaaagca ggaataaaat taacaagatg 841 taattgacat aagtcccatc accgttgtat aaatgtgtgg aattgtgagc ggataacaat 901 ttcacacacc aactcataaa gtcaagtagg agattaattc catggatacc ctcctcattc 961 tcggcgatag tctctccgcc ggttatcgca tgtctgcctc tgccgcctgg cccgccctcc 1021 tcaatgataa atggcaaagc aaaacctctg tggtgaatgc cagtatttcc ggcgatacct 1081 ctcaacaagg cctcgcccgt ctccccgccc tcctcaaaca acaccaaccc cgttgggtgt 1141 tggtggaact cggcggtaat gatggcctcc gcggttttca accccaacaa accgaacaaa 1201 ccctccggca aattctccaa gatgtgaaag ccgccaatgc cgaacccctc ctcatgcaaa 1261 ttcgcctccc cgccaattat ggtcgccgct acaatgaagc ctttagtgcc atttatccca 1321 aactcgccaa agaatttgat gtgcccctcc tccccttttt catggaagaa gtgtacctca 1381 aaccccaatg gatgcaagat gatggtattc accccaatcg ggatgcccaa ccctttattg 1441 ccgattggat ggccaaacaa ctccaacccc tcgtgaatca tgatagttaa tctagatttc 1501 ttgtacaaac tcggccctat ccggctgtct aacaaagccc gaaaggaagc tgagttggct 1561 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgacg 1621 ggttttttgt ctagatcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt 1681 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcattta cagctagctc 1741 agtcctaggt acaatgctag ccggaggagg gtcaattcat ggtggtgaat gtgaaaccag 1801 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg 1861 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg 1921 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga 1981 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta 2041 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg 2101 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca 2161 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc 2221 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg 2281 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc 2341 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat 2401 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgacttt agtgccatgt 2461 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg 2521 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg 2581 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc 2641 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct 2701 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg 2761 tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 2821 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac 2881 gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat gcccttgaga 2941 attaaggagt ggacaacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt 3001 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga 3061 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata 3121 atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt 3181 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac 3241 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg 3301 atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag 3361 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc 3421 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg 3481 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg 3541 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt 3601 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa 3661 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc 3721 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg 3781 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct 3841 aatgacttgg ttcacgtaga ttaagcgatc cgaatggcgg aatcttcggg tattgatgct 3901 accctgaaaa tattttgtag atagggcccc tttagcccac tgatttaaca aaaccgagtt 3961 acatttctct aggaaaacat tcacatggac ggaaaccaag tgcctactgt taccctcaaa 4021 cgcccaatta atctgaaggt catcgtgacc ccccgctgga aagaggaagt acagcagcaa 4081 ctccagggac aaatagcgac ttttgatggt caattacaac aactggatgt ccaaggaaat 4141 cgcaccatta gtgaacttca acaacagggc aatacccccc aggtcgctca acaaatggaa 4201 aatatccaac tccaggttaa ccaaaaaaag cgggaactcc ttgagaagaa aaaccaagtg 4261 ctccagcagc tccaacaggt acaaaactta gaattagatc aagagtttct ccagggtcaa 4321 atggagagtt tctttgaagt gaaagtgggc gataatctgg tagagtcgac ctgcaggcat 4381 gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 4441 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 4501 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 4561 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 4621 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 4681 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 4741 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 4801 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 4861 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 4921 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 4981 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 5041 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 5101 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 5161 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 5221 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 5281 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 5341 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 5401 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 5461 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 5521 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 5581 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 5641 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 5701 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 5761 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 5821 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 5881 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 5941 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 6001 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 6061 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 6121 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 6181 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 6241 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 6301 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 6361 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 6421 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 6481 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 6541 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 6601 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 6661 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 6721 cgtcagggcg cgtca //