LOCUS       sequence-291856-        6735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KO of PCC 7002 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 6735)
  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 6735)
  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: pSZT025
FEATURES             Location/Qualifiers
     source          1..6735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     promoter        1727..1761
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     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"
     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(join(4449..4455,4456..4473,4474..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(join(5562..6353,6354..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
//
