LOCUS       Exported                5275 bp ds-DNA     circular SYN 20-SEP-2019
DEFINITION  Ligation of sfGFP  into PNW33N+Ppta+3Tter.
ACCESSION   .
VERSION     .
KEYWORDS    pNW-sfGFP(SPS6)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5275)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Sep 20, 2019 from SnapGene 4.3.11
            https://www.snapgene.com
COMMENT     Vector molecule:  PNW33N
               Fragment ends: SphI and XbaI
               Fragment size: 4510
            Insert molecule:  sfGFP*
               Fragment ends: XbaI and SphI
               Fragment size: 765
FEATURES             Location/Qualifiers
     source          1..5275
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     9..36
                     /label=Fp_mut_F
                     /note="from pKB01_ sfGFP(Sp)"
     misc_feature    23..27
                     /label='RBS
     misc_feature    34..747
                     /gene="sfGFP(Sp)"
                     /product="optimized by Jan Willem Veening (gfp_sf_JWV)"
                     /label=sfGFP(SPS6)
                     /note="optimized by Jan Willem Veening (gfp_sf_JWV)"
                     /note="/translation=""MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDA
                     TDGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQER
                     TISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITAD
                     KQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEK
                     RDHMVLLEFVTAAGITHGMDELYK"""
     misc_feature    34..36
                     /label=START
     primer_bind     complement(748..775)
                     /label=FP MUT _R
                     /note="from pKB01_ sfGFP(Sp)"
     misc_feature    776..952
                     /label=3TER
                     /note="3fold terminator"
     misc_feature    776..864
                     /label=TER_rrnB
     misc_feature    869..920
                     /label=TER_rpsI
     primer_bind     complement(874..900)
                     /label=GFP_infu_F
                     /note="from s102 into PNW33N"
     misc_feature    925..952
                     /label=TER_tufA
     primer_bind     complement(939..957)
                     /label=GFP_XhoIR
                     /note="from s102 into PNW33N"
     primer_bind     complement(977..997)
                     /label=PNW33NmcsRev
                     /note="sequencing of MCS inserts pNW33n Hao Wai PhD thesis"
     primer_bind     complement(1156..1176)
                     /label=pNW33N_rev
                     /note="sequencing primer pNW33N"
     misc_feature    complement(1215..1865)
                     /label=cat
                     /note="chloramphenicol acetyltransferase"
     misc_feature    complement(1972..2976)
                     /label=repB
                     /note="RepB"
     primer_bind     complement(3071..3092)
                     /label=pNW33N_for
                     /note="sequencing primer pNW33N"
     primer_bind     4959..4980
                     /label=pNW33N_for
                     /note="sequencing primer pNW33N"
     primer_bind     5076..5096
                     /label=PNW33NmcsFor
                     /note="sequencing MCS pNW33N Han Wai thesis"
     primer_bind     5120..5138
                     /label=GFP_BglIIF
                     /note="from s102 into PNW33N"
     primer_bind     5120..5138
                     /label=GFP_infu_R2
                     /note="from s102 into PNW33N"
     misc_feature    5122..5266
                     /label=Ppta
ORIGIN
        1 ggggatcctc tagaactaat aaggaggaca aacatgtcaa aaggagaaga actttttaca
       61 ggtgtagtac ctatcttggt tgaattggat ggtgatgtta acggtcacaa attttctgta
      121 cgtggtgaag gtgaaggtga tgcaactgac ggtaaattga cacttaaatt catttgtaca
      181 actggaaaac ttcctgttcc ttggcctact cttgttacaa cattgacata tggagtacaa
      241 tgtttttcac gttatcctga tcatatgaaa cgtcacgatt tttttaaatc tgctatgcca
      301 gaaggttatg tacaagaacg tacaatttca tttaaagatg acggaacata taaaacacgt
      361 gctgaagtaa aattcgaagg tgacactctt gttaatcgta tcgaattgaa aggaatcgat
      421 ttcaaagaag atggtaacat tttgggacac aaacttgaat acaacttcaa ctctcataat
      481 gtttatatca cagctgacaa acaaaaaaac ggtattaaag ctaattttaa aattcgtcac
      541 aatgttgaag atggatctgt tcaattggcc gatcattatc aacaaaatac accaatcgga
      601 gacggaccag tattgcttcc agataaccac tacctttcta ctcaatcagt tctttcaaaa
      661 gatcctaacg aaaaacgtga ccatatggta cttcttgaat ttgttacagc agcaggtatc
      721 actcacggta tggacgaact ttataaataa taatgagcac tagtcaaggg catgctagga
      781 cgccgccaag ccagcttaaa cccagctcaa tgagctgggt tttttgtttg ttaaaaatga
      841 agaagaaact gtgaagcgta tttatagcaa agcactcaaa agtttacctt atgggtgctt
      901 ttttcgtgct tttttgaaaa gacaaaaaaa agaaccttgc caagcaagat tcaagcttgg
      961 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1021 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1081 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagccct
     1141 tcaaacttcc caaaggcgag ccctagtgac attagaaaac cgactgtaaa aagtacagtc
     1201 ggcattatct catattataa aagccagtca ttaggcctat ctgacaattc ctgaatagag
     1261 ttcataaaca atcctgcatg ataaccatca caaacagaat gatgtacctg taaagatagc
     1321 ggtaaatata ttgaattacc tttattaatg aattttcctg ctgtaataat gggtagaagg
     1381 taattactat tattattgat atttaagtta aacccagtaa atgaagtcca tggaataata
     1441 gaaagagaaa aagcattttc aggtataggt gttttgggaa acaatttccc cgaaccatta
     1501 tatttctcta catcagaaag gtataaatca taaaactctt tgaagtcatt ctttacagga
     1561 gtccaaatac cagagaatgt tttagataca ccatcaaaaa ttgtataaag tggctctaac
     1621 ttatcccaat aacctaactc tccgtcgcta ttgtaaccag ttctaaaagc tgtatttgag
     1681 tttatcaccc ttgtcactaa gaaaataaat gcagggtaaa atttatatcc ttcttgtttt
     1741 atgtttcggt ataaaacact aatatcaatt tctgtggtta tactaaaagt cgtttgttgg
     1801 ttcaaataat gattaaatat ctcttttctc ttccaattgt ctaaatcaat tttattaaag
     1861 ttcatttgat atgcctccta aatttttatc taaagtgaat ttaggaggct tacttgtctg
     1921 ctttcttcat tagaatcaat ccttttttaa aagtcaatcc cgtttgttga actactcttt
     1981 aataaaataa tttttccgtt cccaattcca cattgcaata atagaaaatc catcttcatc
     2041 ggctttttcg tcatcatctg tatgaatcaa atcgccttct tctgtgtcat caaggtttaa
     2101 ttttttatgt atttctttta acaaaccacc ataggagatt aaccttttac ggtgtaaacc
     2161 ttcctccaaa tcagacaaac gtttcaaatt cttttcttca tcatcggtca taaaatccgt
     2221 atcctttaca ggatattttg cagtttcgtc aattgccgat tgtatatccg atttatattt
     2281 atttttcggt cgaatcattt gaacttttac atttggatca tagtctaatt tcattgcctt
     2341 tttccaaaat tgaatccatt gtttttgatt cacgtagttt tctgtattct taaaataagt
     2401 tggttccaca cataccaata catgcatgtg ctgattataa gaattatctt tattatttat
     2461 tgtcacttcc gttgcacgca taaaaccaac aagattttta ttaatttttt tatattgcat
     2521 cattcggcga aatccttgag ccatatctga caaactctta tttaattctt cgccatcata
     2581 aacattttta actgttaatg tgagaaacaa ccaacgaact gttggctttt gtttaataac
     2641 ttcagcaaca accttttgtg actgaatgcc atgtttcatt gctctcctcc agttgcacat
     2701 tggacaaagc ctggatttac aaaaccacac tcgatacaac tttctttcgc ctgtttcacg
     2761 attttgttta tactctaata tttcagcaca atcttttact ctttcagcct ttttaaattc
     2821 aagaatatgc agaagttcaa agtaatcaac attagcgatt ttcttttctc tccatggtct
     2881 cacttttcca ctttttgtct tgtccactaa aacccttgat ttttcatctg aataaatgct
     2941 actattagga cacataatat taaaagaaac ccccatctat ttagttattt gtttggtcac
     3001 ttataacttt aacagatggg gtttttctgt gcaaccaatt ttaagggttt tcaatacttt
     3061 aaaacacata cataccaaca cttcaacgca cctttcagca actaaaataa aaatgacgtt
     3121 atttctatat gtatcaagaa tagaaagaac tcgtttttcg ctacgctcaa aacgcaaaaa
     3181 aagcactcat tcgagtgctt tttcttatcg ctccaaatca tgcgattttt tcctctttgc
     3241 ttttctttgc tcacgaagtt ctcgatcacg ctgcaaaaca tcttgaagcg aaaaagtatt
     3301 cttcttttct tccgatcgct catgctgacg cacgaaaagc cctctaggcg cataggaaca
     3361 actcctaaat gcatgtgagg ggttttctcg tccatgtgaa cagtcgcata cgcaatattt
     3421 tgtttcccat actgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3481 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3541 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3601 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3661 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3721 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3781 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3841 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3901 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3961 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4021 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4081 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4141 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4201 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4261 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4321 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4381 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4441 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4501 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4561 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagcccgat
     4621 atgggaaaca aaatattgcg tatgcgactg ttcacatgga cgagaaaacc cctcacatgc
     4681 atttaggagt tgttcctatg cgcctagagg gcttttcgtg cgtcagcatg agcgatcgga
     4741 agaaaagaag aatacttttt cgcttcaaga tgttttgcag cgtgatcgag aacttcgtga
     4801 gcaaagaaaa gcaaagagga aaaaatcgca tgatttggag cgataagaaa aagcactcga
     4861 atgagtgctt tttttgcgtt ttgagcgtag cgaaaaacga gttctttcta ttcttgatac
     4921 atatagaaat aacgtcattt ttattttagt tgctgaaagg tgcgttgaag tgttggtatg
     4981 tatgtgattc aataatttct tttactcgct cgttatagtc gatcggttca tcattcacca
     5041 aatcataatt ttcatgtgac cgttctttat caatatcggg attcgtttta ctttcccgtt
     5101 ctctctgatt gtgaaattga attcaacgct cgaatgccgc atgtaaaaga taataaactg
     5161 aattgtctta ataaatcgat tttttatggc aatgcttaac ttagttgcgt ttctatataa
     5221 taagagtatc ctagtatata ggggcatttc aaacattagg gaggattcgt tcccc
//
