LOCUS       sequence-271133-        5266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCC 11901 fadD locus KO with a kanamycin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5266)
  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 5266)
  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: pSW071
FEATURES             Location/Qualifiers
     source          1..5266
                     /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"
     primer_bind     complement(1315..1334)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1326..2135
                     /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(1393..1412)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(3045..3061)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3045..3061)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3058..3080)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3069..3085
                     /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(3093..3099,3100..3117,3118..3123))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3138..3159
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3276..3293)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3447..4035)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3527..3546)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4206..4997,4998..5066))
                     /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     4829..4848
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5067..5171)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5239..5257
                     /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 gaattgccac attgcacaag
     1261 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1321 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     1381 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1441 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1501 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1561 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1621 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1681 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1741 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1801 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1861 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     1921 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     1981 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     2041 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     2101 tgcagtttca tttgatgctc gatgagtttt tctaagcgat ccgaatggcg gaatcttcgg
     2161 gtattgatgc taccctgaaa atattttgca gatagggccc ctttagccca ctgatttaac
     2221 aaaaccgagt tacatttctc taggaaaaca ttcacatgga cggaaaccaa gtgcctactg
     2281 ttaccctcaa acgcccaatt aatctgaagg tcatcgtgac cccccgctgg aaagaggaag
     2341 tacagcagca actccaggga caaatagcga cttttgatgg tcaattacaa caactggatg
     2401 tccaaggaaa tcgcaccatt agtgaacttc aacaacaggg caataccccc caggtcgctc
     2461 aacaaatgga aaatattcaa ctccaggtta accaaaaaaa gcgggaactc cttgagaaga
     2521 aaaaccaagt gctccagcag ctccaacagg tacaaaactt agaattagat caagagtttc
     2581 tccagggtca gatggagagt ttctttgaag tgaaagtggg cgataatctg gtgcgcaagc
     2641 tgaatgttga acttgtggtc cgggatggtg ttgttgaaga aattcgcggc gacctctaat
     2701 ttttttgtga ttgctgtcgc gtgacagtgt tcggctcccc cgatgacact tgcggtgcgg
     2761 gggattttta atggggaaaa aattcatgaa aattggcatt gtcgggctgg gtttaatcgg
     2821 tggatctttg gcgatcgcct tccgggaaaa aggcttagaa gtgttgggta tttcccggaa
     2881 aaatacaacc tgcgaaacgg ccttgaggaa gggaattgtc gaccaagcca gcacagattt
     2941 ttctcttctc aaggatacgg atgtggtgtt tctctgcacg ccgatcaaag cgattttgcc
     3001 cacggctcaa acccttattc cccacctcaa atccacggcg atttgtcata gctgtttcct
     3061 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3121 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3181 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     3241 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     3301 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3361 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3421 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3481 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3541 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3601 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3661 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3721 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3781 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3841 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3901 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3961 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4021 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4081 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4141 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4201 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4261 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4321 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4381 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4441 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4501 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4561 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4621 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4681 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     4741 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4801 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4861 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4921 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4981 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5041 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5101 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5161 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     5221 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
