LOCUS       40924_969        6338 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY B helper, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6338)
  AUTHORS   Collier R, Thomson JG, Thilmony R.
  TITLE     GAANTRY, a versatile and robust Agrobacterium-based gene stacking 
            system generates high quality transgenic plants
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6338)
  AUTHORS   Thilmony R.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-2017) Crop Improvement and Genetics research Unit,
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   3  (bases 1 to 6338)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6338)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-DEC-2017) Crop Improvement and Genetics research Unit, 
            USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /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"
     primer_bind     633..649
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    669..690
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        705..735
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    743..759
                     /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)."
     CDS             779..1447
                     /codon_start=1
                     /gene="ParA"
                     /product="ParA"
                     /label=ParA
                     /note="small serine site-specific recombinase"
                     /protein_id="AWK49527.1"
                     /translation="MATREQQPEGRRLKVARIYLRASTDEQNLERQESLVAATRAAGYY
                     VAGIYREKASGARADRPELLRMIADLQPGEVVVAEKIDRISRLPLAEAERLVASIRAKG
                     AKLAVPGVVDLSELAAEANGVAKIVLESVQDMLLKLALQMARDDYEDRRERQRQGVQLA
                     KAAGRYTGRKRDAGMHDRIIALRSGGSSIAKTAKLVGCSPSQVKRVWAAWNAQQQNKRA
                     "
     gene            779..1447
                     /gene="ParA"
                     /label=ParA
     CDS             1455..2813
                     /codon_start=1
                     /gene="A118"
                     /product="A118"
                     /label=A118
                     /note="A118 large serine site-specific recombinase"
                     /protein_id="AWK49528.1"
                     /translation="MKAAIYIRVSTQEQVENYSIQAQTEKLTALCRSKDWDVYDIFIDG
                     GYSGSNMNRPALNEMLSKLHEIDAVVVYRLDRLSRSQRDTITLIEEYFLKNNVEFVSLS
                     ETLDTSSPFGRAMIGILSVFAQLERETIRDRMVMGKIKRIEAGLPLTTAKGRTFGYDVI
                     DTKLYINEEEAKQLQLIYDIFEEEQSITFLQKRLKKLGFKVRTYNRYNNWLTNDLYCGY
                     VSYKDKVHVKGIHEPIISEEQFYRVQEIFTRMGKNPNMNRDSASLLNNLVVCSKCGLGF
                     VHRRKDTMSRGKKYHYRYYSCKTYKHTHELEKCGNKIWRADKLEELIINRVNNYSFASR
                     NVDKEDELDSLNEKLKIEHAKKKRLFDLYINGSYEVSELDSMMNDIDAQINYYESQIEA
                     NEELKKNKKIQENLADLATVDFDSLEFREKQLYLKSLINKIYIDGEQVTIEWL"
     gene            1455..2813
                     /gene="A118"
                     /label=A118
     regulatory      2820..2868
                     /label=T3 transcriptional terminator
                     /note="T3 transcriptional terminator"
                     /regulatory_class="terminator"
     promoter        2888..2992
                     /label=AmpR promoter
     CDS             3002..4081
                     /label=lacI
                     /note="lac repressor"
     terminator      4084..4131
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4189..4205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4213..4229)
                     /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(4237..4267)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4282..4303)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4591..5179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5353..6210)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6211..6315)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgtattggg atgtaaaacg acggccagtg aattgggccc
      661 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
      721 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagccat
      781 ggcaacgcgc gaacagcaac cggagggccg ccgtcttaaa gtggcccgca tttacctgcg
      841 cgcgtcgacg gacgagcaga atcttgaacg ccaggaatcc ctggtcgcgg ccacccgcgc
      901 tgccggctat tatgtcgccg gcatttatcg cgagaaggcc agcggcgcac gggccgaccg
      961 gcctgagctg ctgcgcatga ttgccgatct ccaaccgggg gaagtcgtgg tcgccgaaaa
     1021 aatcgatcgc atcagccggc tcccgctcgc ggaagccgaa cgtctggtcg cctcgatccg
     1081 cgcgaagggt gcgaagctcg cggtgccggg tgtcgtcgat ctgagcgaac tcgcagcgga
     1141 agccaacggc gtcgcgaaga tcgtcctgga atccgtccag gatatgctgc tgaagctggc
     1201 cctgcagatg gctcgtgatg attacgagga ccgccgcgag cgccagcggc agggcgtcca
     1261 gcttgcaaag gctgcgggtc gctatacggg tcgtaagcgc gacgccggca tgcatgaccg
     1321 cattatcgcc ctccgttcgg gcggctcgtc catcgccaag accgcgaaac tggttgggtg
     1381 ctcgccgtcc caggtcaagc gcgtctgggc tgcgtggaac gcccagcagc agaacaagcg
     1441 cgcgtgaggt acccatgaaa gcggcgatct atattcgggt gagcacgcag gagcaggtcg
     1501 aaaattattc cattcaggcg caaacggaga aactgaccgc cctttgccgc tcgaaagatt
     1561 gggatgtgta tgatatcttc atcgacggcg gctactccgg ctcgaacatg aaccgcccgg
     1621 cgctgaatga aatgctgagc aagctgcacg agatcgacgc ggtggttgtg tatcgcctcg
     1681 accgcctgtc ccgctcccag cgggatacga tcacactgat cgaagaatat tttctcaaga
     1741 ataacgtcga gttcgtctcc ctgagcgaaa cgctcgacac cagcagcccg ttcggccgcg
     1801 cgatgatcgg tatcctgtcg gtctttgccc aactggaacg cgaaacgatc cgcgaccgca
     1861 tggtcatggg caagatcaaa cgcatcgaag ccggccttcc gcttacgacg gcgaagggcc
     1921 ggacgttcgg ctacgacgtt attgatacga agctgtacat caacgaggaa gaggccaagc
     1981 agcttcagct gatctacgac atcttcgaag aagaacagtc gatcaccttt cttcagaagc
     2041 gcctgaagaa actgggcttc aaggtgcgca cgtacaaccg gtacaacaac tggcttacga
     2101 atgacctgta ttgcggctac gtgagctata aggataaagt gcatgtgaag ggtatccacg
     2161 agcccatcat ctccgaagag cagttctatc gtgtgcagga aatctttacc cgcatgggca
     2221 agaacccgaa catgaaccgt gatagcgcgt cgctcctcaa caatctggtc gtctgctcca
     2281 agtgcggcct gggtttcgtt caccgccgca aggatacgat gtcgcgcggc aagaagtatc
     2341 attatcgtta ttattcctgt aagacgtata agcataccca tgaactggaa aagtgcggca
     2401 acaagatctg gcgggccgat aaactggagg aactcatcat caaccgcgtc aataactatt
     2461 ccttcgcgtc gcggaatgtg gacaaggagg atgagctgga ttcccttaat gaaaaactga
     2521 agattgagca cgccaagaag aagcgcctct tcgatcttta tatcaacggt agctacgagg
     2581 tctcggaact ggattccatg atgaatgata tcgacgccca gatcaactat tatgaatccc
     2641 agatcgaagc caacgaggag ctgaaaaaga ataaaaagat tcaggagaat cttgcagacc
     2701 tggccacggt ggatttcgac agcctggagt ttcgcgagaa gcagctttac cttaagagcc
     2761 tgatcaataa aatttacatt gatggtgaac aggtgaccat tgaatggctt taagacgtct
     2821 aagcaaaccc cttgggttcc ctctttagga gtctgagggg ttttttgcag tactactagt
     2881 cacgtggcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2941 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggtgaa
     3001 cgtgaagccg gttacgctgt atgacgtggc ggaatatgcg ggcgtgtcgt atcagacggt
     3061 ttcgcgcgtg gtcaaccagg cgtcgcatgt gtcggcgaag acccgcgaaa aggtcgaagc
     3121 ggcaatggcg gaactgaact atatcccgaa ccgcgtcgcc caacagctcg ctggcaagca
     3181 gtccctcctg atcggcgtgg ccacctccag ccttgcgctg catgcgccgt cgcagatcgt
     3241 cgctgccatc aagagccgtg ccgatcaact tggcgcgagc gttgtggttt cgatggtgga
     3301 gcggtcgggc gtcgaagcat gcaaggccgc agtgcataat ctgctcgccc agcgcgtgtc
     3361 cggcctgatc atcaactatc cgcttgatga ccaggacgct attgcagtcg aggccgcatg
     3421 caccaatgtg ccggcgctgt tcctcgatgt ctccgaccag acccccatca actccatcat
     3481 ttttagccat gaagatggca cgcggctggg cgttgaacac ctggtcgctc tgggccacca
     3541 acagatcgcc cttctcgccg gccctctgtc cagcgtgtcc gcccggctcc gtctcgccgg
     3601 ctggcataag tatctcaccc ggaaccagat ccagccgatc gctgaacgcg aaggtgattg
     3661 gtccgcgatg agcggcttcc agcagaccat gcagatgctc aatgaaggca tcgttccgac
     3721 cgcaatgctc gtcgccaacg accaaatggc cctcggcgcc atgcgtgcga tcaccgagtc
     3781 cggccttcgc gtcggcgcag atatctcggt cgtcggttat gatgacacag aagactcctc
     3841 gtgttacatc cctccgtcca cgacgatcaa acaggatttt cgcctgctcg gccagacctc
     3901 cgttgatcgc ctgctgcagc tgtcccaggg tcaggcggtg aaaggtaacc agctgctgcc
     3961 ggtgtccctg gtgaagcgca agaccacgct tgcgccgaac acccagaccg caagcccccg
     4021 cgcacttgcg gattccctga tgcaactggc tcggcaggtc tcgcgtctgg aaagcggcca
     4081 gtgatagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gcggccgcag
     4141 cttggctcga gcatggtcaa tcccaatggc gcgcttggcg taatcatggt catagctgtt
     4201 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4261 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4321 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4381 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4441 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4501 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5281 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5341 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5401 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5461 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5521 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5581 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5641 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5701 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5761 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5821 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5881 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5941 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6001 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6061 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6121 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6181 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6241 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6301 aataggggtt ccgcgcacat ttccccgaaa agtgccac
//
