LOCUS       62056_19610        5332 bp DNA     circular SYN 22-DEC-2019
DEFINITION  Expression vector pSfgfp-Sp, complete sequence.
ACCESSION   MK948098
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5332)
  AUTHORS   Zhang J-Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-MAY-2019) Center for Algal Biology and Applied 
            Research, Institute of Hydrobiology, Donghu Nanlu, Wuhan, Hubei 
            430072, 086
REFERENCE   2  (bases 1 to 5332)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (16-MAY-2019) Center for Algal Biology and Applied Research, 
            Institute of Hydrobiology, Donghu Nanlu, Wuhan, Hubei 430072, 086"
FEATURES             Location/Qualifiers
     source          1..5332
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /label=lac operator
                     /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     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2308..2334
                     /label=protein domain linker
                     /note="protein domain linker"
     CDS             2335..2352
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             2377..3084
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     misc_feature    3085..3114
                     /label=protein domain linker
                     /note="protein domain linker"
     CDS             3121..3144
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             3181..3204
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     terminator      3215..3262
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             3699..4487
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     regulatory      4798..4816
                     /regulatory_class="terminator"
     primer_bind     complement(4938..4954)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctc gcttgcacgc ctctgcactc gtcggtcccg gcatccgatt
     2281 cccgggagta ctggatccaa ggcctctggt ggatctggag gtagtggtgg tagtcaccac
     2341 caccaccacc acggtagcgg tagcggttct gctcgcaaag gtgaagaatt atttactggt
     2401 gttgtaccaa ttctagttga attagatggt gatgttaatg gtcacaaatt ttctgtccgt
     2461 ggtgaaggtg aaggtgatgc tacaaacggt aaattaacct taaaatttat ttgcactact
     2521 ggtaaactac ctgttccttg gccaacatta gtcactactt taacctatgg tgttcaatgc
     2581 ttttcccgtt atcctgatca catgaaacgg catgactttt tcaaaagtgc tatgcccgaa
     2641 ggttatgtac aagaacgcac tatttctttc aaagatgacg gtacatacaa gacccgtgct
     2701 gaagtcaaat ttgaaggtga taccttggtt aatcgtatcg aattaaaagg tattgatttt
     2761 aaagaagatg gtaacattct aggccacaaa ttagaataca actttaactc acacaatgta
     2821 tacatcacag cagataaaca aaagaatggt atcaaagcta acttcaaaat tcgccacaac
     2881 gttgaagatg gttccgttca actagcagac cattatcaac aaaatactcc aattggcgat
     2941 ggccctgtct tattaccaga caaccattac ttgtctacac aatctgtatt gtctaaagat
     3001 cccaacgaaa aacgtgacca catggtttta ttggaatttg taactgctgc tggtattaca
     3061 catggcatgg atgaactata caaaggttct ggtggtggta gcactagcgt cggtagcgct
     3121 tggagccatc cccaatttga aaaaggtggt ggcagcggcg gtggtagcgg tggtagcgct
     3181 tggtctcacc cacaattcga aaagtaagca ataactagca taaccccttg gggcctctaa
     3241 acgggtcttg aggggttttt tgctgaagat gcaccttgcc gtagaagaac agcaaggccg
     3301 ccaatgcctg acgatgcgtg gagaccgaaa ccttgcgctc gttcgccagc caggacagaa
     3361 atgcctcgac ttcgctgctg cccaaggttg ccgggtgacg cacaccgtgg aaacggatga
     3421 aggcacgaac ccagtggaca taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta
     3481 tgcgctcacg caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg
     3541 cggttttcat ggcttgttat gactgttttt ttggggtaca gtctatgcct cgggcatcca
     3601 agcagcaagc gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta
     3661 cgcagcaggg cagtcgccct aaaacaaagt taaacatcat gagggaagcg gtgatcgccg
     3721 aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt
     3781 tgctggccgt acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata
     3841 ttgatttgct ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca
     3901 acgacctttt ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag
     3961 tcaccattgt tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc
     4021 aatttggaga atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg
     4081 acattgatct ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc
     4141 cagcggcgga ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg
     4201 aaaccttaac gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc
     4261 ttacgttgtc ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg
     4321 ctgccgactg ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta
     4381 gacaggctta tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag
     4441 aatttgtcca ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa tgtctaacaa
     4501 ttcgttcaag ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga tgcactaagc
     4561 acataattgc tcacagccaa actatcaggt caagtctgct tttattattt ttaagcgtgc
     4621 ataataagcc ctacacaaat tgggagatat atcatgaaag gctggctttt tcttgttatc
     4681 gcaatagttg gcgaagtaat cgcaacatcc gcattaaaat ctagcgaggg ctttactaag
     4741 ctgatccggt ggatgacctt ttgaatgacc tttaatagat tatattacta attaattggg
     4801 gaccctagag gtcccctttt ttattttaaa aattttttca caaaacggtt tacaagcata
     4861 aagcttgctc aatcaatcac cggatcatca gtactcccgg gatctcgacg cattcgcgaa
     4921 gtaccgatct ccaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4981 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     5041 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     5101 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5161 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5221 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5281 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc ga
//
