LOCUS       Exported                5266 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  PCC 11901 fadD locus KO with a kanamycin resistance cassette.
ACCESSION   .
VERSION     .
KEYWORDS    pSW071
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5266)
  TITLE     Unprecedented biomass and fatty acid production by the newly 
            discovered cyanobacterium Synechococcus sp. PCC 11901
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5266)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5266
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(28..616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(108..127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(787..1647)
                     /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     1410..1429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1648..1752)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1820..1838
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1876..1898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1998..2017
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2211..2233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2225..2242
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2226..2242
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3162..3181)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3173..3982
                     /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(3240..3259)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(4892..4908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4892..4908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4905..4927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4916..4932
                     /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(4940..4970)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4985..5006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5123..5140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
       61 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      121 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      181 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      241 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      301 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      361 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      421 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      481 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
      541 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
      601 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
      661 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
      721 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
      781 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
      841 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
      901 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
      961 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1021 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1081 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1141 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1201 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1261 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1321 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1381 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1441 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1501 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1561 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1621 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1681 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1741 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1801 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     1861 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     1921 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     1981 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     2041 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     2101 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     2161 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     2221 acgttgtaaa acgacggcca gttaattaaa atttgactgc gatcgcctgc aatggcctgg
     2281 gcctgggcga caattttcgg gctaggttcg taatctttgg gggtagcgat gcgaatattc
     2341 atgccgacta acgcgccacc gaggaggatc gagtgggcga cattattgcc atcacccaca
     2401 taggtcatgg tcaggccttc gagttggccg aaggtttcct gcatcgtttg cagatccgcc
     2461 aacatctgac aggggtgttc taaatctgta agggcgttga taatcggcat ttcacagtaa
     2521 tcagcgaagg tttcgagatc ctgttgatca aaggtacgaa tcgccaaaat atcgagatag
     2581 cggtctaaaa ccctggcggt gtcctcgatg ggctcgccgc gacccacttg ggtacggttt
     2641 gggttcaggt caatgacatt gccacccagt tggtacatgg ccacgctaaa ggagactcgg
     2701 gtgcgggtag aggctttgta aaataacaga ccgagggttt tggggcaggt gggggcgatt
     2761 tttccctgtt tgagatctgc cgccaggttt agcagggtaa taatttctgt gctgctgaga
     2821 tctgcaatgc tcagaagatc ccgacccttg agtgcattca tcctgttttc ggctccttac
     2881 cgtggcaaaa aaatcattat aagctcccct ccagattcca caaaagcttg caagggcgat
     2941 cgccactggt cttcgtcagg aaatgttgat ggcgatccca agggggtgac acttgaaaaa
     3001 ctgtctactt ttggggagat gggtgaaatc tggttttaaa gtaggattgt agccatgatt
     3061 tcggcttgtt ttttgcgctt ttaaatggaa ttgccacatt gcacaagata aaaatatatc
     3121 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3181 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     3241 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     3301 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     3361 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     3421 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     3481 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     3541 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     3601 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     3661 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     3721 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     3781 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     3841 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     3901 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     3961 gatgctcgat gagtttttct aagcgatccg aatggcggaa tcttcgggta ttgatgctac
     4021 cctgaaaata ttttgcagat agggcccctt tagcccactg atttaacaaa accgagttac
     4081 atttctctag gaaaacattc acatggacgg aaaccaagtg cctactgtta ccctcaaacg
     4141 cccaattaat ctgaaggtca tcgtgacccc ccgctggaaa gaggaagtac agcagcaact
     4201 ccagggacaa atagcgactt ttgatggtca attacaacaa ctggatgtcc aaggaaatcg
     4261 caccattagt gaacttcaac aacagggcaa taccccccag gtcgctcaac aaatggaaaa
     4321 tattcaactc caggttaacc aaaaaaagcg ggaactcctt gagaagaaaa accaagtgct
     4381 ccagcagctc caacaggtac aaaacttaga attagatcaa gagtttctcc agggtcagat
     4441 ggagagtttc tttgaagtga aagtgggcga taatctggtg cgcaagctga atgttgaact
     4501 tgtggtccgg gatggtgttg ttgaagaaat tcgcggcgac ctctaatttt tttgtgattg
     4561 ctgtcgcgtg acagtgttcg gctcccccga tgacacttgc ggtgcggggg atttttaatg
     4621 gggaaaaaat tcatgaaaat tggcattgtc gggctgggtt taatcggtgg atctttggcg
     4681 atcgccttcc gggaaaaagg cttagaagtg ttgggtattt cccggaaaaa tacaacctgc
     4741 gaaacggcct tgaggaaggg aattgtcgac caagccagca cagatttttc tcttctcaag
     4801 gatacggatg tggtgtttct ctgcacgccg atcaaagcga ttttgcccac ggctcaaacc
     4861 cttattcccc acctcaaatc cacggcgatt tgtcatagct gtttcctgtg tgaaattgtt
     4921 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4981 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaa
//
