LOCUS       62056_19805        6569 bp DNA     circular SYN 21-APR-2020
DEFINITION  Cloning vector pSimple-tub+eGFP, complete sequence.
ACCESSION   MN496120
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6569)
  AUTHORS   Zhang Y, Wang H, Yang R, Wang L, Yang G, Liu T.
  TITLE     Genetic Transformation of Tribonema minus, a Eukaryotic Filamentous 
            Oleaginous Yellow-Green Alga
  JOURNAL   Int J Mol Sci 21 (6), E2106 (2020)
  PUBMED    32204356
REFERENCE   2  (bases 1 to 6569)
  AUTHORS   Zhang Y, Wang H, Liu T.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-SEP-2019) Qingdao Institude of Bioenergy and 
            Bioprocess Technology, Chinese Academy of Sciences, No. 189 Songling
            Road, Laoshan District, Qingdao, Shandong 266101, China
REFERENCE   3  (bases 1 to 6569)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Int J Mol 
            Sci"; date: "2020"; volume: "21"; issue: "6"; pages: "E2106"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-SEP-2019) Qingdao Institude of Bioenergy and Bioprocess 
            Technology, Chinese Academy of Sciences, No. 189 Songling Road, 
            Laoshan District, Qingdao, Shandong 266101, China"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: DNAMAN v. 7.0.2.176
            Sequencing Technology :: 454
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      245..1650
                     /label=derived from Tribonema minus tubulin
                     /note="derived from Tribonema minus tubulin"
                     /regulatory_class="promoter"
     CDS             1651..2367
                     /label=EGFP
                     /note="enhanced GFP"
     primer_bind     complement(2948..2964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3003..3021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3028..3044)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3185..3613
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             3957..4748
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4769..5626
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5800..6388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctgc
      241 ccttggcgat tgagctgccc ttgattccag tgcagtcatc tgctgcgggc gaggcacgcc
      301 tagcatacgt gacccgtaca tatgtcatgt gcaggaagag cgggggaggc gtagtgtggc
      361 ggcatttcga gacgtcgctc ccgtaggtgc ttgctgaacg cgatggtttc aatttttgtg
      421 ccaagatgcg gcagcagcgt tgcagcatgt cgtgcgctca gcagcgaggc gtaccatgat
      481 caagccggcc atgcacgttt ctgtcattgg cccttgcaac gatgacacgt gtcttgtagc
      541 aactgtacac agtcaaattt gtaatggata gcacactgca cattgctgag tctgcaagtt
      601 gtctgtcgtg attctgttgc tgttcgcttt tgagggctca ctgcatgctg gccagcacag
      661 gcaaggcagt atgcaatgct gactgcaact tgtgcctatc aatgggtcaa tctgcaaggg
      721 ggtttgcatt cacacacatt ctctgctttg gtctgtcaag caaatgatca tcaagcaaat
      781 gactacctat tgttcgcatt cagagccgat ccgaaggcaa cgcataatcc gcatgagatc
      841 gcgctcctcc actgcctgct gcctctttca acaccatcga cgtgagatat gtagacctac
      901 tgctcaaaag gacgactcga cttagcacgg ccagcaccaa tctactgcgg aaaaacaacg
      961 gcaatgttcc gcgtgcttgc cgcaacagcc agtctgcacc ggcatgccat catcaggaac
     1021 atcatgaatc atatggcccc ttcacggctt caccacgcat gcacacaaga aaatactccc
     1081 cagggcacgt gctgctggct ctgaaaacag cgccaaatgt acacaggtgt ctgtgggtta
     1141 cttgtccaaa aaaccgttcg cttttttatt tctcgcaatt tgattggcca gcgcaactac
     1201 tgactttcga gggcaaaatt tgaaactcct agcgccgcca agctggtgca caaatcacac
     1261 ttccgaactc gtcctgaccg aagcgtcaag acttgctcgg taccagcgcc agctaagcag
     1321 ctagcagcaa ccaaacctct ccttttcctt taaggtacgt acccggcgcg gctggagagc
     1381 gttgtcaaaa gtcagttggt ggttttgatt ttcacccgct ttctggagct tgggggcatg
     1441 gggcgcgcac gctggcgagg cagctaggca gcgggtcgtg gcgctggtga tttctttctg
     1501 ctgcagatcg tcataccagc cagcgcccct gtacagcttg cactggttgc ccaaacagat
     1561 cgaaactgcc gcccattctc accagccacc tcgtgtgctc tcgtctcttg caggaccact
     1621 tctccagtca acggtaccga gctcgccacc atggtaagca agggcgagga gctgttcacc
     1681 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     1741 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     1801 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     1861 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     1921 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     1981 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     2041 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     2101 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     2161 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     2221 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     2281 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     2341 actctcggca tggacgagct gtacaagtaa ggatccgcgc actgttgacg tgtgcacttc
     2401 aacttgccgt tgaagtgcgg gcttcgcctt gacgcttggg ggggggtcgt gtatttctct
     2461 cgtgcgaggt ggctttctct ctttcgtatg taagagtcgg tggcgcacgg gcagcctttg
     2521 tcaatacgcg cggtgaagtg tacgtttgtt gggggggggc tgggggtagg agtgtgccgc
     2581 accgcaacag gtgtgatgcc atgccctcag ctcctctcgg ctttcgtatg tgattggccc
     2641 atgtcaatgg attaccgcgt aacctactgc aaggttctgg tatgattgtg tgtgctgcct
     2701 tgctgttttc ctttttttgt tatatgcaca tttggatgcc ccctgtctcg gcagagcata
     2761 atcattttga tggcattctt tacttgtgaa tgctcagagt gtgctcttct gcaagtctgt
     2821 gttcgacagt aggtgctgcc ttgctgtttc ccttgttctg tgcagtggtg tgccctgtct
     2881 aggctgcgca gaatcatttt gatgacattc tttacttgct cgagaagggc agcttggcgt
     2941 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctaagggca gcttcaattc
     3001 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
     3061 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     3121 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     3181 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3241 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3301 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3361 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3421 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3481 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3541 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3601 tttaacgcga attttaacaa aattcagggc gcaagggctg ctaaaggaag cggaacacgt
     3661 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     3721 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     3781 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     3841 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     3901 tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg tttcgcatga
     3961 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4021 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4081 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4141 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4201 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4261 tcctgtcatc ccaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4321 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4381 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4441 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4501 aggatctcgt cgtgacccac ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4561 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4621 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4681 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4741 agttcttctg aattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4801 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4861 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4921 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4981 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     5041 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5101 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5161 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5221 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5281 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5341 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5401 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5461 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5521 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5581 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5641 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5701 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5761 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5821 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5881 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5941 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6001 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6061 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6121 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6181 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6241 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6301 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6361 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6421 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6481 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6541 cagcgagtca gtgagcgagg aagcggaag
//
