LOCUS       sequence-271131-        7480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KO of PCC 11901 fadD gene/expresses codon-optimized 'tesA gene from 
            E. coli from inducible cLac143 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7480)
  AUTHORS   Wlodarczyk A, Selao TT, Norling B, Nixon PJ
  TITLE     Newly discovered Synechococcus sp. PCC 11901 is a robust 
            cyanobacterial strain for high biomass production.
  JOURNAL   Commun Biol. 2020 May 7;3(1):215. doi: 10.1038/s42003-020-0910-8.
  PUBMED    32382027
REFERENCE   2  (bases 1 to 7480)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2020-05-7"; volume: "3(1):215. doi"; pages: " 
            10.1038/s42003-020-0910-8"
COMMENT     Sequence Label: pSW068
FEATURES             Location/Qualifiers
     source          1..7480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1267..1286
                     /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    1371..1387
                     /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(2056..2074)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2070..2117
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2215..2249
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     CDS             2279..3361
                     /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(2298..2317)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     complement(3529..3548)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3540..4349
                     /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(3607..3626)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(5259..5275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5259..5275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    5283..5299
                     /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(join(5307..5313,5314..5331,5332..5337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5352..5373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5490..5507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5661..6249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5741..5760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6420..7211,7212..7280))
                     /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     7043..7062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7281..7385)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7453..7471
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagttaatt aaaatttgac tgcgatcgcc
      421 tgcaatggcc tgggcctggg cgacaatttt cgggctaggt tcgtaatctt tgggggtagc
      481 gatgcgaata ttcatgccga ctaacgcgcc accgaggagg atcgagtggg cgacattatt
      541 gccatcaccc acataggtca tggtcaggcc ttcgagttgg ccgaaggttt cctgcatcgt
      601 ttgcagatcc gccaacatct gacaggggtg ttctaaatct gtaagggcgt tgataatcgg
      661 catttcacag taatcagcga aggtttcgag atcctgttga tcaaaggtac gaatcgccaa
      721 aatatcgaga tagcggtcta aaaccctggc ggtgtcctcg atgggctcgc cgcgacccac
      781 ttgggtacgg tttgggttca ggtcaatgac attgccaccc agttggtaca tggccacgct
      841 aaaggagact cgggtgcggg tagaggcttt gtaaaataac agaccgaggg ttttggggca
      901 ggtgggggcg atttttccct gtttgagatc tgccgccagg tttagcaggg taataatttc
      961 tgtgctgctg agatctgcaa tgctcagaag atcccgaccc ttgagtgcat tcatcctgtt
     1021 ttcggctcct taccgtggca aaaaaatcat tataagctcc cctccagatt ccacaaaagc
     1081 ttgcaagggc gatcgccact ggtcttcgtc aggaaatgtt gatggcgatc ccaagggggt
     1141 gacacttgaa aaactgtcta cttttgggga gatgggtgaa atctggtttt aaagtaggat
     1201 tgtagccatg atttcggctt gttttttgcg cttttaaatg gaattgcctc tggacatctc
     1261 caaacgaatt gtgagcgctc acaattcgga attcttaaca aaaaagcagg aataaaatta
     1321 acaagatgta attgacataa gtcccatcac cgttgtataa atgtgtggaa ttgtgagcgg
     1381 ataacaattt cacacaccaa ctcataaagt caagtaggag attaattcca tggataccct
     1441 cctcattctc ggcgatagtc tctccgccgg ttatcgcatg tctgcctctg ccgcctggcc
     1501 cgccctcctc aatgataaat ggcaaagcaa aacctctgtg gtgaatgcca gtatttccgg
     1561 cgatacctct caacaaggcc tcgcccgtct ccccgccctc ctcaaacaac accaaccccg
     1621 ttgggtgttg gtggaactcg gcggtaatga tggcctccgc ggttttcaac cccaacaaac
     1681 cgaacaaacc ctccggcaaa ttctccaaga tgtgaaagcc gccaatgccg aacccctcct
     1741 catgcaaatt cgcctccccg ccaattatgg tcgccgctac aatgaagcct ttagtgccat
     1801 ttatcccaaa ctcgccaaag aatttgatgt gcccctcctc ccctttttca tggaagaagt
     1861 gtacctcaaa ccccaatgga tgcaagatga tggtattcac cccaatcggg atgcccaacc
     1921 ctttattgcc gattggatgg ccaaacaact ccaacccctc gtgaatcatg atagttaatc
     1981 tagatttctt gtacaaactc ggccctatcc ggctgtctaa caaagcccga aaggaagctg
     2041 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     2101 tcttgacggg ttttttgtct agatcaacgg cctcaaccta ctactgggct gcttcctaat
     2161 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcatttaca
     2221 gctagctcag tcctaggtac aatgctagcc ggaggagggt caattcatgg tggtgaatgt
     2281 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     2341 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     2401 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     2461 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     2521 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     2581 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     2641 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     2701 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     2761 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca
     2821 aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg
     2881 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactttag
     2941 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
     3001 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
     3061 gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg
     3121 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
     3181 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
     3241 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
     3301 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3361 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc
     3421 ccttgagaaa atgacgagcg aattagtcat ctttctagat taacattgca caagataaaa
     3481 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     3541 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     3601 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     3661 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     3721 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     3781 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     3841 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     3901 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     3961 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     4021 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     4081 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     4141 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     4201 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     4261 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     4321 ttcatttgat gctcgatgag tttttctaag cgatccgaat ggcggaatct tcgggtattg
     4381 atgctaccct gaaaatattt tgcagatagg gcccctttag cccactgatt taacaaaacc
     4441 gagttacatt tctctaggaa aacattcaca tggacggaaa ccaagtgcct actgttaccc
     4501 tcaaacgccc aattaatctg aaggtcatcg tgaccccccg ctggaaagag gaagtacagc
     4561 agcaactcca gggacaaata gcgacttttg atggtcaatt acaacaactg gatgtccaag
     4621 gaaatcgcac cattagtgaa cttcaacaac agggcaatac cccccaggtc gctcaacaaa
     4681 tggaaaatat tcaactccag gttaaccaaa aaaagcggga actccttgag aagaaaaacc
     4741 aagtgctcca gcagctccaa caggtacaaa acttagaatt agatcaagag tttctccagg
     4801 gtcagatgga gagtttcttt gaagtgaaag tgggcgataa tctggtgcgc aagctgaatg
     4861 ttgaacttgt ggtccgggat ggtgttgttg aagaaattcg cggcgacctc taattttttt
     4921 gtgattgctg tcgcgtgaca gtgttcggct cccccgatga cacttgcggt gcgggggatt
     4981 tttaatgggg aaaaaattca tgaaaattgg cattgtcggg ctgggtttaa tcggtggatc
     5041 tttggcgatc gccttccggg aaaaaggctt agaagtgttg ggtatttccc ggaaaaatac
     5101 aacctgcgaa acggccttga ggaagggaat tgtcgaccaa gccagcacag atttttctct
     5161 tctcaaggat acggatgtgg tgtttctctg cacgccgatc aaagcgattt tgcccacggc
     5221 tcaaaccctt attccccacc tcaaatccac ggcgatttgt catagctgtt tcctgtgtga
     5281 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5341 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5401 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5461 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5521 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5581 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5821 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6061 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6361 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6421 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6481 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6541 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6601 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6661 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6721 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6781 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6841 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6901 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6961 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7021 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7081 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7141 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7201 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7261 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7321 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7381 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7441 ttaacctata aaaataggcg tatcacgagg ccctttcgtc
//
