LOCUS       62056_8690        6580 bp DNA     circular SYN 21-APR-2020
DEFINITION  Cloning vector pEASY-tub/neo, complete sequence.
ACCESSION   MN481464
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  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 6580)
  AUTHORS   Zhang Y, Wang H, Liu T.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-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 6580)
  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 
            (19-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..6580
                     /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      253..1625
                     /label=derived from Tribonema minus tubulin
                     /note="derived from Tribonema minus tubulin"
                     /regulatory_class="promoter"
     CDS             1626..2417
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      2421..2974
                     /label=derived from Tribonema minus tubulin
                     /note="derived from Tribonema minus tubulin"
                     /regulatory_class="terminator"
     promoter        complement(3014..3032)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3039..3055)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3196..3624
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             3968..4759
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4780..5637
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5811..6399
                     /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 gccaagcttg
      241 gtaccgagct cgtcggatcc cagtgcagtc atctgctgcg ggcgaggcac gcctagcata
      301 cgtgacccgt acatatgtca tgtgcaggaa gagcggggga ggcgtagtgt ggcggcattt
      361 cgagacgtcg ctcccgtagg tgcttgctga acgcgatggt ttcaattttt gtgccaagat
      421 gcggcagcag cgttgcagca tgtcgtgcgc tcagcagcga ggcgtaccat gatcaagccg
      481 gccatgcacg tttctgtcat tggcccttgc aacgatgaca cgtgtcttgt agcaactgta
      541 cacagtcaaa tttgtaatgg atagcacact gcacattgct gagtctgcaa gttgtctgtc
      601 gtgattctgt tgctgttcgc ttttgagggc tcactgcatg ctggccagca caggcaaggc
      661 agtatgcaat gctgactgca acttgtgcct atcaatgggt caatctgcaa gggggtttgc
      721 attcacacac attctctgct ttggtctgtc aagcaaatga tcatcaagca aatgactacc
      781 tattgttcgc attcagagcc gatccgaagg caacgcataa tccgcatgag atcgcgctcc
      841 tccactgcct gctgcctctt tcaacaccat cgacgtgaga tatgtagacc tactgctcaa
      901 aaggacgact cgacttagca cggccagcac caatctactg cggaaaaaca acggcaatgt
      961 tccgcgtgct tgccgcaaca gccagtctgc accggcatgc catcatcagg aacatcatga
     1021 atcatatggc cccttcacgg cttcaccacg catgcacaca agaaaatact ccccagggca
     1081 cgtgctgctg gctctgaaaa cagcgccaaa tgtacacagg tgtctgtggg ttacttgtcc
     1141 aaaaaaccgt tcgctttttt atttctcgca atttgattgg ccagcgcaac tactgacttt
     1201 cgagggcaaa atttgaaact cctagcgccg ccaagctggt gcacaaatca cacttccgaa
     1261 ctcgtcctga ccgaagcgtc aagacttgct cggtaccagc gccagctaag cagctagcag
     1321 caaccaaacc tctccttttc ctttaaggta cgtacccggc gcggctggag agcgttgtca
     1381 aaagtcagtt ggtggttttg attttcaccc gctttctgga gcttgggggc atggggcgcg
     1441 cacgctggcg aggcagctag gcagcgggtc gtggcgctgg tgatttcttt ctgctgcaga
     1501 tcgtcatacc agccagcgcc cctgtacagc ttgcactggt tgcccaaaca gatcgaaact
     1561 gccgcccatt ctcaccagcc acctcgtgtg ctctcgtctc ttgcaggacc acttctccag
     1621 tcaacatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     1681 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     1741 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     1801 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     1861 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     1921 ggcaggatct cctgtcatcc caccttgctc ctgccgagaa agtatccatc atggctgatg
     1981 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     2041 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     2101 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     2161 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     2221 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     2281 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     2341 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     2401 ttcttgacga gttcttctga ggatccacta gtgcgcactg ttgacgtgtg cacttcaact
     2461 tgccgttgaa gtgcgggctt cgccttgacg cttggggggg ggtcgtgtat ttctctcgtg
     2521 cgaggtggct ttctctcttt cgtatgtaag agtcggtggc gcacgggcag cctttgtcaa
     2581 tacgcgcggt gaagtgtacg tttgttgggg gggggctggg ggtaggagtg tgccgcactg
     2641 caacaggtgt gatgccatgc cctcagctcc tctcggcttt cgtatgtgat tggcccatgt
     2701 caatggatta ccgcgtaacc tactgcaagg ttctggtatg attgtgtgtg ctgccttgct
     2761 gttttccttt ttttgttata tgcacatttg gatgccccct gtctcggcag agcataatca
     2821 ttttgatggc attctttact tgtgaatgct cagagtgtgc tcttctgcaa gtctgtgttc
     2881 gacagtaggt gctgccttgc tgtttccctt gttctgtgca gtggtgtgcc ctgtctaggc
     2941 tgcgcagaat cattttgatg acattcttta cttgctcgag catgcatcta gagggcccaa
     3001 tcgcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     3061 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3121 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3181 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     3241 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     3301 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     3361 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     3421 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     3481 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     3541 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3601 tttaacaaaa atttaacgcg aattttaaca aaattcaggg cgcaagggct gctaaaggaa
     3661 gcggaacacg tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta
     3721 ctgggctatc tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg
     3781 gcttacatgg cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc
     3841 agctggggcg ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt
     3901 gccgccaagg atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc
     3961 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4021 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4081 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4141 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4201 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4261 ggggcaggat ctcctgtcat cccaccttgc tcctgccgag aaagtatcca tcatggctga
     4321 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4381 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4441 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     4501 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4561 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4621 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4681 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4741 ccttcttgac gagttcttct gaattgaaaa aggaagagta tgagtattca acatttccgt
     4801 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     5041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5641 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5701 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5761 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5821 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5881 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5941 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6001 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6061 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6121 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6181 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6241 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6301 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6361 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6421 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     6481 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6541 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
