LOCUS       sequence-291855-        5143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  KO of PCC 11901 acsA gene/expresses YFP under control of the cpt 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5143)
  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 5143)
  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: pSW036
FEATURES             Location/Qualifiers
     source          1..5143
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             267..296
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             309..320
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(342..344,345..347,348..1061)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(387..408)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(648..667)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     995..1016
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1131..1149)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1145..1192
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1945..1961)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1945..1961)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1958..1980)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1969..1985
                     /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(1993..1999,2000..2017,2018..2023))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2038..2059
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2176..2193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2347..2935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2427..2446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3106..3897,3898..3966))
                     /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     3729..3748
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3967..4071)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4139..4157
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4195..4217)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4317..4336
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4530..4552
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4544..4561
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4545..4561
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aggtcatatc cgaggcgtac attcaatcaa cttgagcgca aaaatcttgc aaaccattac
       61 ggcttaagca cgagaccccc tgatggggcg gacttggggg acaatggggt gaaataaatc
      121 ccacgataat aattaacaaa aaagcaggaa taaaattaac aagatgtaac agacataagt
      181 cccatcaccg ttgtataaag ttaactgtgg gattgcaaaa ggagctccca actcataaag
      241 tcaagtagga gattaattcc atgggccatc atcatcatca tcatcatcat catcacagca
      301 gcggccatat cgaaggtcgt catatgcagg cagcagcctc gatggtgagc aagggcgagg
      361 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      421 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      481 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
      541 acggcctgca gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      601 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      661 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      721 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      781 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
      841 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
      901 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtccg
      961 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
     1021 ccgccgggat cactctcggc atggacgagc tgtacaagta acgagggcgg tgctttggca
     1081 ggatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     1141 ataactagca taaccccttg gggcctctaa acgggtcttg acgggttttt tgtctagatc
     1201 aacggcctca aagatgtcca gaggacagag ggtgtcttcc cttgagggga gtcggaggga
     1261 aaatttccat taggtgtccc tccgactgct tgccttgacc tagttccctt cgataaaggc
     1321 gaaacggccc tgttgacccg attcatccat ttcaatttgg gccacaaaaa attccttctg
     1381 gacaatttcc ccttcctctg tgaaggaaat ttcacccaaa ggcgtgacgt aggtacctgc
     1441 aaaaatttcg tcccgcagtt gtcgtcgcaa gtccggtaga gcaagagttt ctaggggcgt
     1501 tttttcgtcg aggctgctga gggcttcaac aaaaacttgg atcgccgtaa aagcctgggc
     1561 actaaattgg ggcggctctt tttggttttg ttgaaaatag gcgtcccgaa acgcgcggtt
     1621 aatctcatta tctaactcgg cactgtaggc ttgggccacc aacaccccat cacattttgc
     1681 ttggcagacg gggaaaatat tagaagtatt taagccgttc cccccaacaa taatgccgtt
     1741 gtaacctaat tctcgcagtt gcttcaccag gttgcccgca tcggctgcta accccgaaat
     1801 aatcactaga tctggttgaa cattgagtgc ggcagtcacc tgggcttgaa aatcagtgtc
     1861 ggtggtttga aacctttgta ccgtgactaa ttccagacca aaatcttgga cagccgtctg
     1921 gaaggtttct gtctctgatt tactgtcata gctgtttcct gtgtgaaatt gttatccgct
     1981 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2041 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2101 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     2161 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     2221 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     2281 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     2341 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2401 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2461 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2521 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2581 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2641 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2701 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2761 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2821 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2881 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2941 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3001 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3061 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3121 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     3181 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     3241 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     3301 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3361 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3421 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3481 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3541 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3601 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3661 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3721 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3781 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3841 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3901 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3961 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4021 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4081 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4141 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4201 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4261 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4321 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     4381 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     4441 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     4501 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     4561 tgcaatgctg agatgatcct cggtgaactc cttgatcgcc aagcctttgg tgtggtgcaa
     4621 ccgttactgt ttgatccagc gatcgccccg gagaatgaaa ccattgcttt tttacgggga
     4681 cgctatatct ggcaaacaac cttggccgcc gtcccaccat cgccaaccca agcaacggcg
     4741 atcgccacgg tgcaagaaaa ttgggacatg gccctagccc aacagcccaa cgaccccgaa
     4801 attttgatgg ccctgggttt tgtccattac ctcaatgcag attacgaagc cgccaatgat
     4861 cgttggtatg aagccttagc agcggcaacg acccaggatt tccccccagt cctccaggcg
     4921 gagatttatg cgggcctggc gattagtatg tacgatttgg cccaagggca acccaccgac
     4981 gttgaccagg aaattcttgt ggggaaagcc gaaaaactcc gggcgatcgc cctagatctc
     5041 gatcgttccc agctcaaccc ccgccgcctc gaacaaagtt ggttatggtt ccctgaggcg
     5101 atcgccactt ggcaaaccct gctcgcttta gaagaatctc cct
//
