LOCUS       40924_34709       10297 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPIPRA721, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10297)
  AUTHORS   Figueroa-Balderas RE, Chi-Ham CL, Le H, Tricoli D, Sandman J, 
            Johnson D, Reich J, Bennett AB.
  TITLE     Constitutive expression of eIF5A3 increases high quality biomass 
            yield in an elite alfalfa cultivar
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10297)
  AUTHORS   Figueroa-Balderas RE, Chi-Ham CL, Le H, Tricoli D, Sandman J, 
            Johnson D, Reich J, Bennett AB.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2015) Plant Sciences, UC Davis, One shields 
            Avenue, Davis, CA 95616, USA
REFERENCE   3  (bases 1 to 10297)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10297)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2015) Plant Sciences, UC Davis, One shields Avenue, Davis, 
            CA 95616, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10297
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..406
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     CDS             420..1142
                     /codon_start=1
                     /gene="ipt"
                     /product="isopentenyl phosphotransferase"
                     /label=ipt
                     /note="involved in condensation of AMP and 
                     isopentenylpyrophosphate to form isopentenyl-AMP"
                     /protein_id="AKE42824.1"
                     /translation="MDLHLIFGPTCTGKTTTAIALAQQTGLPVLSLDRVQCCPQLSTGS
                     GRPTVEELKGTTRLYLDDRPLVEGIIAAKQAHHRLIEEVYNHEANGGLILEGGSTSLLN
                     CMARNSYWSADFRWHIIRHKLPDQETFMKAAKARVKQMLHPAAGHSIIQELVYLWNEPR
                     LRPILKEIDGYRYAMLFASQNQITADMLLQLDANMEGKLINGIAQEYFIHARQQEQKFP
                     QVNAAAFDGFEGHPFGMY"
     gene            420..1142
                     /gene="ipt"
                     /label=ipt
     terminator      1505..1757
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    1782..1949
                     /label=T-DNA left border
                     /note="T-DNA left border"
     regulatory      1975..2556
                     /label=histone H3
                     /note="histone H3"
                     /regulatory_class="promoter"
     CDS             2598..3077
                     /codon_start=1
                     /gene="eIF5A3"
                     /product="eukaryotic translation initiation factor 5A"
                     /label=eIF5A3
                     /note="eIF5A3; derived from Populus deltoides"
                     /protein_id="AKE42825.1"
                     /translation="MSDEEQHFESKADAGASKTYPQQAGTIRKSGYIVIKNRPCKVVEV
                     STSKTGKHGHAKCHFVAIDIFNGKKLEDIVPSSHNCDVPHVTRTDYQLIDISEDGFVSL
                     LTENGNTKDDLRLPTDESLLSQIKDGFGEGKDLVVTVMSSMGEEQICALKDVGPK"
     gene            2598..3077
                     /gene="eIF5A3"
                     /label=eIF5A3
                     /note="PdeIF5A3"
     regulatory      3100..3403
                     /label=pea E9
                     /note="pea E9"
                     /regulatory_class="terminator"
     misc_feature    3434..3642
                     /label=T-DNA left border
                     /note="T-DNA left border"
     CDS             4981..5607
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1
                     (Heeb et al., 2000)"
     CDS             6039..7109
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     rep_origin      7178..7372
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7846..7986
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(8172..8760)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9228..10019)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 tctaggtcgg cggatcgatt gagatttttc aaatcagtgc gcaagacgtg acgtaagtat
       61 ctgagctagt ttttattttt ctactaattt ggtcgtttat ttcggcgtgt aggacatggc
      121 aaccgggcct gaatttcgcg ggtattctgt ttctattcca actttttctt gatccgcagc
      181 cattaacgac ttttgaatag atacgctgac acgccaagcc tcgctagtca aaagtgtacc
      241 aaacaacgct ttacagcaag aacggaatgc gcgtgacgct cgcggtgacg ccatttcgcc
      301 ttttcagaaa tggataaata gccttgcttc ctattatatc ttcccaaatt accaatacat
      361 tacactagca tctgaatttc ataaccaatc tcgatacacc aaatcgaatc gataccggta
      421 tggacctgca tctaattttc ggtccaactt gcacaggaaa gacgacgacc gcgatagctc
      481 ttgcccagca gacagggctt ccagtccttt cgcttgatcg ggtccaatgc tgtcctcaac
      541 tatcaaccgg aagcggacga ccaacagtgg aagaactgaa aggaacgacg cgtctctacc
      601 ttgatgatcg gcctctggtg gagggtatca tcgcagccaa gcaagctcat cataggctga
      661 tcgaggaggt gtataatcat gaggccaacg gcgggcttat tcttgaggga ggatccacct
      721 cgttgctcaa ctgcatggcg cgaaacagct attggagtgc agattttcgt tggcatatta
      781 ttcgccacaa gttacccgac caagagacct tcatgaaagc ggccaaggcc agagttaagc
      841 agatgttgca ccccgctgca ggccattcta ttattcaaga gttggtttat ctttggaatg
      901 aacctcggct gaggcccatt ctgaaagaga tcgatggata tcgatatgcc atgttgtttg
      961 ctagccagaa ccagatcacg gcagatatgc tattgcagct tgacgcaaat atggaaggta
     1021 agttgattaa tgggatcgct caggagtatt tcatccatgc gcgccaacag gaacagaaat
     1081 tcccccaagt taacgcagcc gctttcgacg gattcgaagg tcatccgttc ggaatgtatt
     1141 aggttacgcc agccctgcgt cgcacctgtc ttcatctgga taagatgttc gtaattgttt
     1201 ttggctttgt cctgttgtgg cagggcggca aatacttccg acaatccatc gtgtcttcaa
     1261 actttatgct ggtgaacaag tcttagtttc cacgaaagta ttatgttaaa ttttaaaatt
     1321 tcgatgtata atgtggctat aattgtaaaa ataaactatc gtaagtgtgc gtgttatgta
     1381 taatttgtct aaatgtttaa tatatatcat agaacgcaat aaatattaaa tatagcgctt
     1441 ttatgaaata taaatacatc attacaagtt gtttatattt cgggtggacc ggtgcgatcg
     1501 caatcttgga ctcccatgtt ggcaaaggca accaaacaaa caatgaatga tccgctcctg
     1561 catatggggc ggtttgagta tttcaactgc catttgggct gaattgaaga catgctcctg
     1621 tcagaaattc cgtgatctta ctcaatattc agtaatctcg gccaatatcc taaatgtgcg
     1681 tggctttatc tgtctttgta ttgtttcatc aattcatgta acgtttgctt ttcttatgaa
     1741 ttttcaaata aattatcaga tcgcgatcgc cccgatgtac atggctggaa gcttggcggc
     1801 aggatgtata cagaggtata caattttata ttacatttat atttgtgtta attcattgaa
     1861 ttttcacttt tattttttac tttgataatc aactgtgtaa agaattattt gaaaaatata
     1921 tataatttat agaatttttt tttgttatgt gtacatctag taatgcatgg cgccatcgat
     1981 gatatccaaa gctttacatt cagggactat tttgcaattt acctgcattc agggactaaa
     2041 gtgacgactt ctttcctatt cagagactaa agtgaccaat ctctcaaaat gaagatattt
     2101 tgttttgttt tagcaggaat gcgaagagtt aaaggcgaga taagttgcac acgatttaca
     2161 ctcacaaaag aaacacaaat tgtccacgct ggcaatccgc aactttacaa accaaccaat
     2221 cagaaacaaa cacacggatc gcacttaata ttttcactta aaaaactcat cattaccgtt
     2281 gaagcattca aagtccacga tctctctact ctaattaacc ttccttaatc atcattaacc
     2341 cttgcatata taaacacact tctcttcaac aaccctcatt acacatttct tctctttcgc
     2401 taaatctaat caatctttcc ctctcttcga gctttctctc tccgattccc ataggtaacc
     2461 accgttcacc gccgtaacgg ttttttcttc tttctgtttt cttgatctta gggtttcgtt
     2521 ttcttcaatt cgaatttttt gattgatttc atcgattttt tggttcaggc ggccgcactc
     2581 tcttatcaat cgcggccatg tctgacgagg agcagcactt cgagtcaaaa gctgatgcgg
     2641 gagcttcgaa aacttaccct caacaagctg gtaccattcg caagagcggt tacattgtca
     2701 tcaagaatcg cccttgcaag gttgtggagg tttctacctc taaaactggc aagcacggcc
     2761 atgccaaatg tcactttgtt gcaattgata tcttcaatgg aaaaaagctt gaagatattg
     2821 ttccttcttc ccacaactgt gatgttcccc atgtcacccg tactgactat cagctgattg
     2881 atatctcaga ggatggattt gtgagcttgc tgactgagaa tggcaatacc aaggatgacc
     2941 tgaggctccc aactgatgag agtctcctct ctcagatcaa ggatggattt ggcgagggta
     3001 aagatcttgt tgtgactgtg atgtcctcca tgggagagga gcagatctgc gccctcaagg
     3061 acgttggccc gaagtaaatt gggatccccg ggtaccgcga tcgcttcgag tattatggca
     3121 ttgggaaaac tgtttttctt gtaccatttg ttgtgcttgt aatttactgt gttttttatt
     3181 cggttttcgc tatcgaactg tgaaatggaa atggatggag aagagttaat gaatgatatg
     3241 gtccttttgt tcattctcaa attaatatta tttgtttttt ctcttatttg ttgtgtgttg
     3301 aatttgaaat tataagagat atgcaaacat tttgttttga gtaaaaatgt gtcaaatcgt
     3361 ggcctctaat gaccgaagtt aatatgagga gtaaaacatc gcgatcgcgg cgccatgcat
     3421 tactagacaa ttgttatgcg ggctaacggg ctgcccgcgg ccctttcggg ctagccctaa
     3481 cgggtaccgg gccccggcag gatgtataca gaggtataca ctagttgtga ataagtcgct
     3541 gtgtatgttt gtttgagatc tctaagagaa aagagcgttt attagaataa tcggatattt
     3601 aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtcaattgcc tagacatgac
     3661 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     3721 cggtgagcgc taaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3781 agcggatgcc gggagcagat agggcccaac ccctccgctg ctatagtgca gtcggcttct
     3841 gacgttcagt gcagccgtct tctgaaaacg acatgtcgca caagtcctaa gttacgcgac
     3901 aggctgccgc cctgcccttt tcctggcgtt ttcttgtcgc gtgttttagt cgcataaagt
     3961 agaatacttg cgactagaac cggagacatt acgccatgaa caagagcgcc gccgctggcc
     4021 tgctgggcta tgcccgcgtc agcaccgacg accaggactt gaccaaccaa cgggccgaac
     4081 tgcacgcggc cggctgcacc aagctgtttt ccgagaagat caccggcacc aggcgcgacc
     4141 gcccggagct ggccaggatg cttgaccacc tacgccctgg cgacgttgtg acagtgacca
     4201 ggctagaccg cctggcccgc agcacccgcg acctactgga cattgccgag cgcatccagg
     4261 aggccggcgc gggcctgcgt agcctggcag agccgtgggc cgacaccacc acgccggccg
     4321 gccgcatggt gttgaccgtg ttcgccggca ttgccgagtt cgagcgttcc ctaatcatcg
     4381 accgcacccg gagcgggcgc gaggccgcca aggcccgagg cgtgaagttt ggcccccgcc
     4441 ctaccctcac cccggcacag atcgcgcacg cccgcgagct gatcgaccag gaaggccgca
     4501 ccgtgaaaga ggcggctgca ctgcttggcg tgcatcgctc gaccctgtac cgcgcacttg
     4561 agcgcagcga ggaagtgacg cccaccgagg ccaggcggcg cggtgccttc cgtgaggacg
     4621 cattgaccga ggccgacgcc ctggcggccg ccgagaatga acgccaagag gaacaagcat
     4681 gaaaccgcac caggacggcc aggacgaacc gtttttcatt accgaagaga tcgaggcgga
     4741 gatgatcgcg gccgggtacg tgttcgagcc gcccgcgcac gtctcaaccg tgcggctgca
     4801 tgaaatcctg gccggtttgt ctgatgccaa gctggcggcc tggccggcca gcttggccgc
     4861 tgaagaaacc gagcgccgcc gtctaaaaag gtgatgtgta tttgagtaaa acagcttgcg
     4921 tcatgcggtc gctgcgtata tgatgcgatg agtaaataaa caaatacgca aggggaacgc
     4981 atgaaggtta tcgctgtact taaccagaaa ggcgggtcag gcaagacgac catcgcaacc
     5041 catctagccc gcgccctgca actcgccggg gccgatgttc tgttagtcga ttccgatccc
     5101 cagggcagtg cccgcgattg ggcggccgtg cgggaagatc aaccgctaac cgttgtcggc
     5161 atcgaccgcc cgacgattga ccgcgacgtg aaggccatcg gccggcgcga cttcgtagtg
     5221 atcgacggag cgccccaggc ggcggacttg gctgtgtccg cgatcaaggc agccgacttc
     5281 gtgctgattc cggtgcagcc aagcccttac gacatatggg ccaccgccga cctggtggag
     5341 ctggttaagc agcgcattga ggtcacggat ggaaggctac aagcggcctt tgtcgtgtcg
     5401 cgggcgatca aaggcacgcg catcggcggt gaggttgccg aggcgctggc cgggtacgag
     5461 ctgcccattc ttgagtcccg tatcacgcag cgcgtgagct acccaggcac tgccgccgcc
     5521 ggcacaaccg ttcttgaatc agaacccgag ggcgacgctg cccgcgaggt ccaggcgctg
     5581 gccgctgaaa ttaaatcaaa actcatttga gttaatgagg taaagagaaa atgagcaaaa
     5641 gcacaaacac gctaagtgcc ggccgtccga gcgcacgcag cagcaaggct gcaacgttgg
     5701 ccagcctggc agacacgcca gccatgaagc gggtcaactt tcagttgccg gcggaggatc
     5761 acaccaagct gaagatgtac gcggtacgcc aaggcaagac cattaccgag ctgctatctg
     5821 aatacatcgc gcagctacca gagtaaatga gcaaatgaat aaatgagtag atgaatttta
     5881 gcggctaaag gaggcggcat ggaaaatcaa gaacaaccag gcaccgacgc cgtggaatgc
     5941 cccatgtgtg gaggaacggg cggttggcca ggcgtaagcg gctgggttgt ctgccggccc
     6001 tgcaatggca ctggaacccc caagcccgag gaatcggcgt gagcggtcgc aaaccatccg
     6061 gcccggtaca aatcggcgcg gcgctgggtg atgacctggt ggagaagttg aaggccgcgc
     6121 aggccgccca gcggcaacgc atcgaggcag aagcacgccc cggtgaatcg tggcaagcgg
     6181 ccgctgatcg aatccgcaaa gaatcccggc aaccgccggc agccggtgcg ccgtcgatta
     6241 ggaagccgcc caagggcgac gagcaaccag attttttcgt tccgatgctc tatgacgtgg
     6301 gcacccgcga tagtcgcagc atcatggacg tggccgtttt ccgtctgtcg aagcgtgacc
     6361 gacgagctgg cgaggtgatc cgctacgagc ttccagacgg gcacgtagag gtttccgcag
     6421 ggccggccgg catggccagt gtgtgggatt acgacctggt actgatggcg gtttcccatc
     6481 taaccgaatc catgaaccga taccgggaag ggaagggaga caagcccggc cgcgtgttcc
     6541 gtccacacgt tgcggacgta ctcaagttct gccggcgagc cgatggcgga aagcagaaag
     6601 acgacctggt agaaacctgc attcggttaa acaccacgca cgttgccatg cagcgtacga
     6661 agaaggccaa gaacggccgc ctggtgacgg tatccgaggg tgaagccttg attagccgct
     6721 acaagatcgt aaagagcgaa accgggcggc cggagtacat cgagatcgag ctagctgatt
     6781 ggatgtaccg cgagatcaca gaaggcaaga acccggacgt gctgacggtt caccccgatt
     6841 actttttgat cgatcccggc atcggccgtt ttctctaccg cctggcacgc cgcgccgcag
     6901 gcaaggcaga agccagatgg ttgttcaaga cgatctacga acgcagtggc agcgccggag
     6961 agttcaagaa gttctgtttc accgtgcgca agctgatcgg gtcaaatgac ctgccggagt
     7021 acgatttgaa ggaggaggcg gggcaggctg gcccgatcct agtcatgcgc taccgcaacc
     7081 tgatcgaggg cgaagcatcc gccggttcct aatgtacgga gcagatgcta gggcaaattg
     7141 ccctagcagg ggaaaaaggt cgaaaaggtc tctttcctgt ggatagcacg tacattggga
     7201 acccaaagcc gtacattggg aaccggaacc cgtacattgg gaacccaaag ccgtacattg
     7261 ggaaccggtc acacatgtaa gtgactgata taaaagagaa aaaaggcgat ttttccgcct
     7321 aaaactcttt aaaacttatt aaaactctta aaacccgcct ggcctgtgca taactgtctg
     7381 gccagcgcac agccgaagag ctgcaaaaag cgcctaccct tcggtcgctg cgctccctac
     7441 gccccgccgc ttcgcgtcgg cctatcgcgg ccgctggccg ctcaaaaatg gctggcctac
     7501 ggccaggcaa tctaccaggg cgcggacaag ccgcgccgtc gccactcgac cgccggcgct
     7561 gggcccttat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7621 tatttagaaa aaccatggta aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     7681 accttaaacc attattatga cgtcctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     7741 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     7801 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     7861 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     7921 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     7981 atcaggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     8041 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     8101 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     8161 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     8221 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     8281 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     8341 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     8401 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     8461 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     8521 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     8581 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctc cgctctgctg
     8641 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     8701 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     8761 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     8821 gggattttgg tcatggacgt cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     8881 gcggcgagcg gtatcagctc actcaaaggc ggtaggcgcg ccatcgatgg cacgaaccca
     8941 gtggacataa gcctcgttcg gttcgtaagc tgtaatgcaa gtagcgtaac tgccgtcacg
     9001 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     9061 ggcttcttgt tatgacatgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
     9121 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
     9181 agggcagtcg ccctaaaaca aagttaaaca tcatcgatac cggtctaaaa caattcatcc
     9241 agtaaaatat aatattttat tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat
     9301 agctcgacat actgttcttc cccgatatcc tccctgatcg accggacgca gaaggcaatg
     9361 tcataccact tgtccgccct gccgcttctc ccaagatcaa taaagccact tactttgcca
     9421 tctttcacaa agatgttgct gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg
     9481 ggcttttccg tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg agtgtcttct
     9541 tcccagtttt cgcaatccac atcggccaga tcgttattca gtaagtaatc caattcggct
     9601 aagcggctgt ctaagctatt cgtataggga caatccgata tgtcgatgga gtgaaagagc
     9661 ctgatgcact ccgcatacag ctcgataatc ttttcagggc tttgttcatc ttcatactct
     9721 tccgagcaaa ggacgccatc ggcctcactc atgagcagat tgctccagcc atcatgccgt
     9781 tcaaagtgca ggacctttgg aacaggcagc tttccttcca gccatagcat catgtccttt
     9841 tcccgttcca catcataggt ggtcccttta taccggctgt ccgtcatttt taaatatagg
     9901 ttttcatttt ctcccaccag cttatatacc ttagcaggag acattccttc cgtatctttt
     9961 acgcagcggt atttttcgat cagttttttc aattccggtg atattctcat tttagccata
    10021 ccggtgcgat cgctgtctaa caattcgttc aagccgacgc cgcttcgccg gcgttaactc
    10081 aagcgattag atgcactaag cacataattg ctcacagcca aactatcagg tcaagtctgc
    10141 ttttattatt tttaagcgtg cataataagc cctacacaaa ttgggagata tatcatggcg
    10201 atcgcggcgc gcctgactga ctgacatacg gttatccaca gaatcagggg ataacgcagg
    10261 aaagaacatg tgagcaaaag gccagcaaaa ggccagg
//
