LOCUS       sequence-291868-        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"
     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"
     promoter        2415..2449
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     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"
     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(join(5507..5513,5514..5531,5532..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(join(6620..7411,7412..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
//
