LOCUS       62056_12835        6272 bp DNA     circular SYN 08-APR-2020
DEFINITION  Expression vector pIgNAT, complete sequence.
ACCESSION   MK903009
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6272)
  AUTHORS   Balestreri C, Stern RF, Wheeler G, Brownlee C, Highfield A.
  TITLE     pIgNAT, a nourseothricin resistance construct to detect 
            transformation capabilities in the alga, Isochryis galbana
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6272)
  AUTHORS   Faktorova D, Nisbet E, Jose FR, Casacuberta E, Allen AE.
  TITLE     Genetic tool development in marine protists: Emerging model 
            organisms for experimental cell biology
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 6272)
  AUTHORS   Balestreri C, Stern RF, Wheeler G, Brownlee C, Highfield A.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-MAY-2019) CPR survey, Marine Biological Association, 
            Citadel Hill, Plymouth PL1 2PB, United Kingdom
REFERENCE   4  (bases 1 to 6272)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (08-MAY-2019) CPR survey, Marine Biological Association, Citadel 
            Hill, Plymouth PL1 2PB, United Kingdom"
FEATURES             Location/Qualifiers
     source          1..6272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     3'UTR           complement(402..1560)
                     /gene="Hsp70"
                     /label=derived from Emiliania huxleyi
                     /note="derived from Emiliania huxleyi"
     CDS             complement(1564..2124)
                     /label=NrsR
                     /note="nourseothricin acetyltransferase"
     5'UTR           complement(2137..4002)
                     /gene="Hsp70"
                     /label=derived from Emiliania huxleyi
                     /note="derived from Emiliania huxleyi"
     primer_bind     complement(4051..4067)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4075..4091)
                     /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)."
     promoter        complement(4099..4129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4144..4165)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4453..5041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5215..6072)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6073..6177)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cctcgtcgag tgtggcgcgt
      421 ggcggcggcc tctgggcggc gcgcctcgga gccctcccgg agccatgtcg gtccctcccc
      481 taggcgcggg gagcggcctc tgggcggcgc tgctccgcag gcgtggcgtc gccgtcgccg
      541 cctacgacgc cgtgccgtgg gtgtgcgagg ctggctttgg cgaggtgcag cgcggcggcc
      601 ccgagcgggt gggccgccat gcggacgcgt ctctgctgct ctgctggccg ccgctcgagg
      661 cctccgcctc cggcagcaac acgatgggcg tccgagcgct gcgcgccatg acgggcagag
      721 gcgcgctccg cccgagctgt ggttgccggc cgcgcggcac gcctcacgcc ccgctcacgc
      781 gccctccccc gccgcggcgc cacacctcag gcgcgcttct gtacgtcggc gagtggcgcg
      841 gcgccgtcgg cgtcctctct tcgctggcgc cttccaccgc cgaccacggg atggttggcg
      901 ggccgcagct gcaggccgag gtggagcggc actggcggct cgccgagtcc gtcgccctgc
      961 cccgctggcc aggcgtgggg agcccgcagg cggggcaccg cctctacgtc tttactcgcc
     1021 gccagccacc gcccgagccc gcggaggagg atgcagcgcc gccgcccccg ccgccccccc
     1081 gccgccgccg ccgccgccgc caccgccgcc gcctccgccg ccgcctccgc cgccgcctcc
     1141 gccgccgccg ccgcctccgc cgcccactca gcgacagccg ccgcccgttt cggatggcta
     1201 cgccgcccgg cgcgagaggc tctctcggct gcagggtgaa gtggacgcgc ctggccggga
     1261 gtgggcgtgg ctcatcgcca gcctgctgtg ggaagaagct tgccgcccag aggagatttg
     1321 atgaaatgaa aagtaatata tctttacttc catgttccaa cacccggacg cgcgttccca
     1381 ggcgccaggg tgatggaagg ttcgacgcac ccccggactg ggcggagaaa tgaaacattt
     1441 cggaaaacgg tgtggtccta tacacccgcc cacaccgcga cacccctcgc acgcggtcat
     1501 ctccgctcgg tgcatatccc tagggtggga gcttctcctc tgaatgcact ctgagccgaa
     1561 tcaggggcag ggcatcgaca tgtagagggc ctgctcgccg tccgaggccg tgccgtcgta
     1621 gagcgccgtg tcgaggccgc agagggtgaa gcccatgcga cggtaggcgt ggatggccgg
     1681 ggcgttgacg ttcgtgacct cgagccagag gtggccagcg ccgcgctcgc gcgcgaactc
     1741 ggtggcgagg cccatgagcg cgcggccgac gccgtggccg cggtgctcgg gggcgacctc
     1801 gatgtcctcg acggtgaggc gacggttcca gcccgagtac gagacgacga cgaagccggc
     1861 gaggtcgccg tcgtcgccgt aggcgacgaa cgtgcgcgag tccgggtcgc cgtcctcgcc
     1921 ggcgtcggac tcgtcgtccg actcgtcgtc ggggaagacc ttcgtgaggg gcgggtcgac
     1981 ggggacctcg cggagcgtga agccgtcgcc cgtggcggtg acgcggaaga cggtgtccgt
     2041 ggtgaacgag ccgtcgagcg cctcgatggc ctccgcgtcg ccgggcaccg acgtgcggta
     2101 gcggtacgcc gtgtcgtcga gcgtcgccgc cgccatggtg tccaagagcg cctactcgcg
     2161 tgacggcgct gcgtcacagg cagggggtca acaggggcta ggcgccaccg cgggcgcacg
     2221 tctcaagtcg agagccttgc acaacacgtg ttcgaggctc gcgacaaaaa atacccctgc
     2281 tcctcaaatc cttgctgggt cgctgtccct tgtcaggagc aggagaacta ggagagcgca
     2341 accagtagga ggcccgcgcc tgcccccgtg ccgccgggtg caacctgtcc tatacgagag
     2401 aggaatcccg gaatccctgg gaccctgtcg tgtgggtgaa ccctacccag cccccctccg
     2461 tgtcgcgatg cgctccccgc tcccgttccc ctgtgtccct gatcgtcctt ctccaagatc
     2521 ctgctcgtgc aggtggggac cttgtcctcc tcccatccct cgtttggttt gtctgttctc
     2581 tctctctctc tctgcacctt ggccaatgtt tagcgtttag gcccaaaagc ctctcgaaag
     2641 gacagaccaa gcaagtccgt ggcactggca agaggtgaag agccacatcg gcgaacacac
     2701 ctgcttggag cacttggagt gcagtcggta acctcacatc cagggccgac ctctgctgtt
     2761 gctttcatct cgcgcgatct gcctcttgcg tcttgggtct tgccaagagc caagaggcaa
     2821 gaccttgaag tggtcttgcg taaggagatt ggccgcccga tagtgcggtc tgtgtgctgg
     2881 ggccttttct agatttctac atctctagct actcgcgtga cggcgctgcg tcacaggcag
     2941 ggggtcaaca ggggctaggc gccaccgcgg gcgcacgtct caagtcgaga gccttgcaca
     3001 acacgtgttc gaggctcgcg acaaaaaata ccttcctcct cctcaaatcc ttgctgggtc
     3061 gctgtccctt gtcaggagca ggagaactag gagagcgcaa ccagtaggag gcccgcgcct
     3121 gcccccgtgc cgccgagtgc aacctgtcct acacgagaga ggaatcccgg aatccctgtg
     3181 ggaccctgtc gtgtgtgtgg accctgtttc ttctgtttca gcccccctcc gtgtcgcgat
     3241 gcgctccccg ctcccgttcc cctgtgtccc tgatcgtcct tctccaagat cctgctcgtg
     3301 caggtgggga ccttgtcctc caggcccaaa agcctctcga aacacgaaag gacagaccac
     3361 agcaagtctg gactggggca ctggcaagag ccatgcgaac acctgctctt gcgagccttg
     3421 gagcacttgg agcagccttg cttgtttttt ttcacctcta taactcctgt ccacgtggcg
     3481 tggtgaggga ctccggaaac aaagcccatg catgggcttt caggagtcat gggctttccg
     3541 gagtccctca ccacgcatgt gcatgtgcat gtgcatgtgc atgtgcatgt gcatgcagcc
     3601 tccctcctca aaggcgtgcg tggacgccgc cgcttgccca tttttgaacg ccgtgaagaa
     3661 gttaactttg gtcatcatgc gtcatggatg gacactgata taagtcagcc ccggtcggga
     3721 atttcttcct ctgaccgaaa ggaacgtttc taaattcttc ctttgcccga gagacgctag
     3781 aactcgtccg ttccccgcga cgtgcgtccc agcgagcgag acataacaaa gcaatgagcc
     3841 atcgcagcga gaggagcgac aagagcgatg gcacgagcgc gccgccgccg cgtcccgccg
     3901 cgccacaggc gccgtctctg atgtcgaggc tgaaccacgg cacgacgagg tcggggagca
     3961 acgtacggag cagcgtgctg agcgtgcgtg tcccctcacc tcggatcctc tagagtcgac
     4021 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4081 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     4141 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4201 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4261 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4321 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4381 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4441 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4501 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4561 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4621 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4681 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4741 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4801 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4861 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4921 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4981 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5041 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5101 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5161 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5221 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5281 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5341 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5401 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5461 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5521 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5581 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     5641 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5701 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5761 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5821 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5881 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5941 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6001 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6061 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6121 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6181 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     6241 taaaaatagg cgtatcacga ggccctttcg tc
//
