LOCUS       40924_999        6419 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector GAANTRY P helper, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6419)
  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  JOURNAL   Unpublished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 6419)
  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 6419)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6419)
  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            "Unpublished"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..6419
                     /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="AWK49531.1"
                     /translation="MATREQQPEGRRLKVARIYLRASTDEQNLERQESLVAATRAAGYY
                     VAGIYREKASGARADRPELLRMIADLQPGEVVVAEKIDRISRLPLAEAERLVASIRAKG
                     AKLAVPGVVDLSELAAEANGVAKIVLESVQDMLLKLALQMARDDYEDRRERQRQGVQLA
                     KAAGRYTGRKRDAGMHDRIIALRSGGSSIAKTAKLVGCSPSQVKRVWAAWNAQQQNKRA
                     "
     gene            779..1447
                     /gene="ParA"
                     /label=ParA
     CDS             1449..2906
                     /codon_start=1
                     /gene="TP901-1"
                     /product="TP901-1"
                     /label=TP901-1
                     /note="large serine site-specific recombinase"
                     /protein_id="AWK49532.1"
                     /translation="MTKKVAIYTRVSTTNQAEEGFSIDEQIDRLTKYAEAMGWQVSDTY
                     TDAGFSGAKLERPAMQRLINDIENKAFDTVLVYKLDRLSRSVRDTLYLVKDVFTKNKID
                     FISLNESIDTSSAMGSLFLTILSAINEFERENIKERMTMGKLGRAKSGKSMMWTKTAFG
                     YYHNRKTGILEIVPLQATIVEQIFTDYLSGISLTKLRDKLNESGHIGKDIPWSYRTLRQ
                     TLDNPVYCGYIKFKDSLFEGMHKPIIPYETYLKVQKELEERQQQTYERNNNPRPFQAKY
                     MLSGMARCGYCGAPLKIVLGHKRKDGSRTMKYHCANRFPRKTKGITVYNDNKKCDSGTY
                     DLSNLENTVIDNLIGFQENNDSLLKIINGNNQPILDTSSFKKQISQIDKKIQKNSDLYL
                     NDFITMDELKDRTDSLQAEKKLLKAKISENKFNDSTDVFELVKTQLGSIPINELSYDNK
                     KKIVNNLVSKVDVTADNVDIIFKFQLA"
     gene            1449..2906
                     /gene="TP901-1"
                     /label=TP901-1
     regulatory      2907..2955
                     /label=T3 transcriptional terminator
                     /note="T3 transcriptional terminator"
                     /regulatory_class="terminator"
     promoter        2969..3073
                     /label=AmpR promoter
     CDS             3083..4162
                     /label=lacI
                     /note="lac repressor"
     terminator      4165..4212
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4270..4286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4294..4310)
                     /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(4318..4348)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4363..4384)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4672..5260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5434..6291)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6292..6396)
                     /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 cgcgtgacat gacaaagaag gtggccatct acacgcgcgt ttccacgacc aaccaggcgg
     1501 aagaaggctt ttcgattgac gaacagatcg atcgccttac gaaatacgcg gaagcaatgg
     1561 gttggcaggt ctccgacaca tacacggatg cgggtttttc gggtgccaaa ctggagcgcc
     1621 cggccatgca gcggctgatc aatgacattg aaaacaaggc ctttgacaca gtgctggtct
     1681 ataaactgga ccgcctctcg cgcagcgtcc gggatacgct gtacctcgtc aaggatgtgt
     1741 ttaccaaaaa taagatcgac ttcattagcc tcaacgaatc gatcgatacc agcagcgcta
     1801 tggggtccct ctttctcacc atcctctcgg ctatcaacga atttgaacgg gagaacatca
     1861 aggagcgtat gacgatgggc aagctcggcc gtgccaagtc cggcaaatcg atgatgtgga
     1921 cgaagaccgc cttcggctac tatcataatc gcaagaccgg catccttgag atcgtcccgc
     1981 tgcaggcaac catcgtggaa cagatcttta ccgactacct gagcggcatc tccctgacca
     2041 agctccgtga caagctgaat gagagcggcc acatcggcaa ggacatcccg tggtcctatc
     2101 gcaccctccg ccagacgctg gacaacccgg tctactgtgg ctatatcaag ttcaaggact
     2161 cgctgtttga agggatgcat aagccgatca tcccgtatga gacatatctc aaggtgcaga
     2221 aggagctgga agagcgccaa cagcagacgt atgaacgcaa taacaacccg cgccccttcc
     2281 aggccaaata tatgctctcg ggcatggctc ggtgcggcta ctgcggcgcc ccgctgaaga
     2341 tcgtgctcgg ccacaagcgc aaggatggct cccgcaccat gaagtatcac tgcgccaacc
     2401 gcttcccccg caagaccaag ggcattacag tttataatga caacaagaaa tgcgactccg
     2461 gcacctatga cctctccaac ctggagaaca cggtcattga caacctgatc ggcttccagg
     2521 aaaacaacga tagcctgctg aaaatcatca atggtaacaa ccagccgatc cttgatacgt
     2581 cgtcctttaa gaagcagatt agccagatcg ataagaagat ccagaagaat agcgatctgt
     2641 atctgaacga tttcatcacg atggacgagc tgaaggaccg taccgacagc ctccaggccg
     2701 aaaagaaact cctcaaggcc aagatcagcg aaaataagtt caacgactcc accgacgtgt
     2761 ttgagctggt caagacccag ctcggctcga tcccgatcaa tgaactgagc tacgataaca
     2821 aaaaaaagat cgtcaacaac ctggtttcga aggtggatgt caccgccgac aacgtcgata
     2881 tcatcttcaa gttccagctg gcgtaataag caaacccctt gggttccctc tttaggagtc
     2941 tgaggggttt tttgcactag tcacgtggcg cggaacccct atttgtttat ttttctaaat
     3001 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3061 aaaaaggaag agtatggtga acgtgaagcc ggttacgctg tatgacgtgg cggaatatgc
     3121 gggcgtgtcg tatcagacgg tttcgcgcgt ggtcaaccag gcgtcgcatg tgtcggcgaa
     3181 gacccgcgaa aaggtcgaag cggcaatggc ggaactgaac tatatcccga accgcgtcgc
     3241 ccaacagctc gctggcaagc agtccctcct gatcggcgtg gccacctcca gccttgcgct
     3301 gcatgcgccg tcgcagatcg tcgctgccat caagagccgt gccgatcaac ttggcgcgag
     3361 cgttgtggtt tcgatggtgg agcggtcggg cgtcgaagca tgcaaggccg cagtgcataa
     3421 tctgctcgcc cagcgcgtgt ccggcctgat catcaactat ccgcttgatg accaggacgc
     3481 tattgcagtc gaggccgcat gcaccaatgt gccggcgctg ttcctcgatg tctccgacca
     3541 gacccccatc aactccatca tttttagcca tgaagatggc acgcggctgg gcgttgaaca
     3601 cctggtcgct ctgggccacc aacagatcgc ccttctcgcc ggccctctgt ccagcgtgtc
     3661 cgcccggctc cgtctcgccg gctggcataa gtatctcacc cggaaccaga tccagccgat
     3721 cgctgaacgc gaaggtgatt ggtccgcgat gagcggcttc cagcagacca tgcagatgct
     3781 caatgaaggc atcgttccga ccgcaatgct cgtcgccaac gaccaaatgg ccctcggcgc
     3841 catgcgtgcg atcaccgagt ccggccttcg cgtcggcgca gatatctcgg tcgtcggtta
     3901 tgatgacaca gaagactcct cgtgttacat ccctccgtcc acgacgatca aacaggattt
     3961 tcgcctgctc ggccagacct ccgttgatcg cctgctgcag ctgtcccagg gtcaggcggt
     4021 gaaaggtaac cagctgctgc cggtgtccct ggtgaagcgc aagaccacgc ttgcgccgaa
     4081 cacccagacc gcaagccccc gcgcacttgc ggattccctg atgcaactgg ctcggcaggt
     4141 ctcgcgtctg gaaagcggcc agtgatagca taaccccttg gggcctctaa acgggtcttg
     4201 aggggttttt tgcggccgca gcttggctcg agcatggtca atcccaatgg cgcgcttggc
     4261 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4321 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4381 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4441 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5101 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5341 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5401 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5461 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5521 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5581 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5641 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5701 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5761 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5821 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5881 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5941 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6001 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6061 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6121 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6181 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6241 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6301 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6361 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
