LOCUS       40924_45818        5769 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pUT3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5769)
  AUTHORS   Kasai Y, Matsuzaki K, Ikeda F, Yoshimitsu Y, Harayama S.
  TITLE     Precise excision of a selectable marker gene in transgenic 
            Pseudococcomyxa strains by the piggyBac transposase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5769)
  AUTHORS   Kasai Y, Harayama S.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-JUN-2017) Contact:Yuki Kasai Chuo University, Research
            and Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 
            112-8551, Japan
REFERENCE   3  (bases 1 to 5769)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5769)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (08-JUN-2017) Contact:Yuki Kasai Chuo University, Research and 
            Development Initiative; 1-13-27 Kasuga, Bunkyo-ku, Tokyo 112-8551, 
            Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5769
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      694..1341
                     /gene="EF1a"
                     /note="Includes promoter region of Pseudococcomyxa sp.
                     strain KJ nuclear gene EF1a for elongation factor 1 alpha"
                     /regulatory_class="promoter"
     gene            694..1341
                     /gene="EF1a"
                     /label=EF1a
     regulatory      1336..1345
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1342..2847
                     /codon_start=1
                     /gene="UMPS"
                     /product="uridine monophosphate synthase"
                     /label=UMPS
                     /protein_id="BAY00744.1"
                     /translation="MATSTPSVATAAELKPKEGPITSQEFEELVLRLHEIEAVKFGNFK
                     LKSGLMSPIYIDLRVIVSYPDVLRRVAEVMWHQVSGAGFDVMCGVPYTALPIATCMSLL
                     HGTPMLMRRKEVKEYGTKKAIEGAFKKGQTCLIVEDLVTSGASVMETVEPLEVEGLKVA
                     DVVVLIDREQGGAARMASNGLRLHSAFTLSFIIKTLQAHGLVSAEVADSVAAFIAANQT
                     FSPSAAAAPTPASAPPAQPKRLPFGERAALCQNAAGRKLLELMARKRTNLAVAADVATV
                     EEMLRIADAAGPHIAVFKTHVDIFDEWDDGIAAQLRHLADKHEFLIFEDRKFADIGNTV
                     VSQYGGGIYKIADWSDITNAHLVPGPGIIDGLRKVGQEKGRGLLLLAEMSSKGALATGA
                     YTEKVAEAAAANQDFVMGFICQSPAKWATSVPPGLVHMTPGVQLASGSDALGQQYNTPA
                     SVIGQGGSDVIIVGRGIIKAADPAAAAAQYREAGWAAYEATLA"
     gene            1342..2847
                     /gene="UMPS"
                     /label=UMPS
     regulatory      2853..3544
                     /gene="EF1a"
                     /note="Includes terminator region of Pseudococcomyxa sp.
                     strain KJ nuclear gene EF1a for elongation factor 1 alpha"
                     /regulatory_class="terminator"
     gene            2853..3544
                     /gene="EF1a"
                     /label=EF1a
     promoter        complement(3581..3599)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3620..3636)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3644..3660)
                     /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(3668..3698)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3713..3734)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4022..4610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4784..5641)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5642..5746)
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgtaatg gccatatcga agcactgttc
      721 tggaaatcta ttcaagtttg aggacaaaaa tgcatcatta gaccgcagtc tgacgtgccc
      781 tgaagcctga gcctgacatg tgatgcctcg tcgtgcaaca aatcgtggcg ggtgaacagc
      841 gctatttaaa agaagagcca gctcaaactg actcctgttt ataggatatt catgataagg
      901 aatcagaatc actttccaca caccaaagtg atctagtact gccgcaagat caaaactcgt
      961 ggaagagtgc aagaccgcgc ttgcaaagaa tcacaatagc ccaatgcttt cagtagcaga
     1021 caactcttaa gtgctaattt gctgcccaca atttccgggg ctggagtttc agagcattcc
     1081 agacgccact tgccagattt ggctccatca gggaactgga acatcgagag agctctctgt
     1141 caagccgcct ctcctgccca ccagcaaaag tatcctttgg catccaacct ttggtacctc
     1201 tgtcacaggt gtggttgctt tccgagccgt ctaacgtctt ttctgtctga aagcgaggtg
     1261 gtttttgtgt tgcgcttgcc acagaaaggc ttggtgcaag tgatctgacc agcttatctg
     1321 tttgcaggaa aactcgccac catggcaact tcaactccct cagtggccac agctgcggag
     1381 ctgaaaccaa aggaagggcc catcacgtca caggagtttg aagagctcgt gctgcgtttg
     1441 catgaaatcg aggccgtcaa atttggcaac ttcaagctca agagcgggct catgtcgccg
     1501 atctacatcg atctgcgcgt cattgtgtcc tatccggacg tcctgcgcag ggtggcagag
     1561 gtgatgtggc atcaggtgtc gggtgctggc tttgacgtga tgtgcggcgt gccatacacg
     1621 gcgctgccca tagcgacatg catgtcgctg ctgcacggga cgcccatgct gatgcggcgc
     1681 aaggaggtga aggagtacgg caccaagaag gccatcgagg gcgccttcaa gaagggccag
     1741 acctgcctca tcgtggagga tctggtcacc agcggcgcct ccgtcatgga gactgtggag
     1801 cccctcgagg tggaggggct gaaggtggcg gatgtggtgg tgctcataga ccgggagcag
     1861 ggcggcgcgg cgcgcatggc cagcaacggg ctgcgactgc actccgcctt cacgctgtcc
     1921 ttcatcatca agacgctgca ggcgcacggc ctcgtcagcg ccgaggtggc cgacagcgtc
     1981 gctgccttca tagccgccaa ccaaaccttc agcccctctg cagcagccgc ccccacaccc
     2041 gcgtctgccc ctcctgcgca gcccaaacgg ctgccatttg gggagcgcgc ggcgctgtgt
     2101 cagaacgcag ccggcaggaa gctgctggag ctgatggcgc gcaagcggac caacctggcg
     2161 gtggcggcgg acgtggcgac ggtggaggag atgctgcgca tcgcggacgc ggccgggccg
     2221 cacatcgccg tcttcaagac ccacgtggac atttttgatg agtgggatga tggcatcgcc
     2281 gcccagctgc gccatctggc cgacaagcat gagttcttga tattcgagga caggaagttt
     2341 gcggacatcg gtaacacagt ggtgtcgcag tacggcggag gcatctacaa gatcgccgac
     2401 tggtcagaca tcaccaatgc acacctggtc ccgggcccag gcatcatcga tggtctccgc
     2461 aaggtgggcc aggagaaggg gcgaggcctg ctgctgctgg ctgagatgag ctccaaggga
     2521 gctcttgcca caggggccta caccgagaag gttgctgaag cggcagctgc caaccaggac
     2581 tttgtcatgg gcttcatctg ccagtcacca gccaagtggg ccacttcagt gcctccaggt
     2641 ctggtgcaca tgacgccggg tgtgcagctg gcgtctggga gcgatgcact ggggcagcag
     2701 tacaacacac ctgcttctgt catcggccag ggcggcagcg atgtcatcat tgtggggaga
     2761 gggatcatca aggcggcaga ccccgctgca gcggccgcac agtacagaga agccggctgg
     2821 gccgcctatg aggccacgct ggcatgacca tggccgctga gagctgcgtg aggatctggg
     2881 gtaggagctg tgcagcctct gccgctgctg gcccctgctg ttgtacacaa gtgccgcccg
     2941 cccagcggga ggtgctcgga ccctgctggc atgccttggt tgctaggcag gctcagaact
     3001 gggtgctgga ctttaggtgg cgtggcttca cgcagctcct tgtctgagca gggcttctgc
     3061 cctgcggcgt gtgcgccttt ctgtgtgcat cccccggatc tctccgttgc agtacttgtc
     3121 agcagtctct cttttccctg tcagtaggat aggccagagc tactccggct tccatcaaac
     3181 attttggaac ttttcatcct gcgcttcttg tcttcacgat cctaggatgt gtggtgggga
     3241 tgatacgctt tgtattcaca cactcgagtt caacaacttt cttagggtgt tcaactgggg
     3301 aggggcattg tgaccatcct gacagacata catctcacac gtcaaagagg gcttgaaaag
     3361 gagtgcatat gcatgtgcat atcctgtggt tgttgagact acaggaagag tgtaaagggt
     3421 tcaactcaca agaaaagttt ctgtggaagc agtgcgtcag tgcatgcatg gactgagtta
     3481 agagccgcag gttgtcacac acttcacatc ctgttgttta gaagcatcat gtttgcaagc
     3541 actcgcggcc gccaccgcgg tggagctcca gcttttgttc cctttagtga gggttaattg
     3601 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     3661 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     3721 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3781 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     3841 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3901 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3961 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4021 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4081 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4141 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4201 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4261 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4321 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4381 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4441 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4501 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4561 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4621 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4681 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4741 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4801 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4861 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4921 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4981 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5041 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5101 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5161 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5221 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5281 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5341 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5401 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5461 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5521 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5581 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5641 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5701 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5761 aagtgccac
//
