LOCUS       62056_3160        7827 bp DNA     circular SYN 25-MAY-2020
DEFINITION  UNVERIFIED: Cloning vector pAPi, complete sequence.
ACCESSION   MK975251
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  AUTHORS   Foley SJ, Orr RG, Galotto G, Liu B, Vidali L.
  TITLE     Robust survival-based RNAi in plants using in tandem silencing of 
            adenine phosphoribosyltransferase
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7827)
  AUTHORS   Foley SJ, Orr RG, Galotto G, Liu B, Vidali L.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-MAY-2019) Biology and Biotechnology, Worcester 
            Polytechnic Institute, 100 Institute Rd., Worcester, MA 01609, USA
REFERENCE   3  (bases 1 to 7827)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-MAY-2019) Biology and Biotechnology, Worcester Polytechnic 
            Institute, 100 Institute Rd., Worcester, MA 01609, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
            GenBank staff is unable to verify sequence and/or annotation 
            provided by the submitter.
FEATURES             Location/Qualifiers
     source          1..7827
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3195..3447
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        3480..3824
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3854..4876
                     /codon_start=1
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4948..5124
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     primer_bind     complement(5681..5697)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5705..5721)
                     /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(5729..5759)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5774..5795)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6083..6671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6845..7702)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7703..7807)
                     /label=AmpR promoter
     promoter        7827
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
ORIGIN
        1 ctagagataa tgagcattgc atgtctaagt tataaaaaat taccacatat tttttttgtc
       61 acacttgttt gaagtgcagt ttatctatct ttatacatat atttaaactt tactctacga
      121 ataatataat ctatagtact acaataatat cagtgtttta gagaatcata taaatgaaca
      181 gttagacatg gtctaaagga caattgagta ttttgacaac aggactctac agttttatct
      241 ttttagtgtg catgtgttct cctttttttt tgcaaatagc ttcacctata taatacttca
      301 tccattttat tagtacatcc atttagggtt tagggttaat ggtttttata gactaatttt
      361 tttagtacat ctattttatt ctattttagc ctctaaatta agaaaactaa aactctattt
      421 tagttttttt atttaataat ttagatataa aatagaataa aataaagtga ctaaaaatta
      481 aacaaatacc ctttaagaaa ttaaaaaaac taaggaaaca tttttcttgt ttcgagtaga
      541 taatgccagc ctgttaaacg ccgtcgacga gtctaacgga caccaaccag cgaaccagca
      601 gcgtcgcgtc gggccaagcg aagcagacgg cacggcatct ctgtcgctgc ctctggaccc
      661 ctctcgagag ttccgctcca ccgttggact tgctccgctg tcggcatcca gaaattgcgt
      721 ggcggagcgg cagacgtgag ccggcacggc aggcggcctc ctcctcctct cacggcacgg
      781 cagctacggg ggattccttt cccaccgctc cttcgctttc ccttcctcgc ccgccgtaat
      841 aaatagacac cccctccaca ccctctttcc ccaacctcgt gttgttcgga gcgcacacac
      901 acacaaccag atctccccca aatccacccg tcggcacctc cgcttcaagg tacgccgctc
      961 gtcctccccc cccccccctc tctaccttct ctagatcggc gttccggtcc atggttaggg
     1021 cccggtagtt ctacttctgt tcatgtttgt gttagatccg tgtttgtgtt agatccgtgc
     1081 tgctagcgtt cgtacacgga tgcgacctgt acgtcagaca cgttctgatt gctaacttgc
     1141 cagtgtttct ctttggggaa tcctgggatg gctctagccg ttccgcagac gggatcgatt
     1201 tcatgatttt ttttgtttcg ttgcataggg tttggtttgc ccttttcctt tatttcaata
     1261 tatgccgtgc acttgtttgt cgggtcatct tttcatgctt ttttttgtct tggttgtgat
     1321 gatgtggtct ggttgggcgg tcgttctaga tcggagtaga attaattctg tttcaaacta
     1381 cctggtggat ttattaattt tggatctgta tgtgtgtgcc atacatattc atagttacga
     1441 attgaagatg atggatggaa atatcgatct aggataggta tacatgttga tgcgggtttt
     1501 actgatgcat atacagagat gctttttgtt cgcttggttg tgatgatgtg gtgtggttgg
     1561 gcggtcgttc attcgttcta gatcggagta gaatactgtt tcaaactacc tggtgtattt
     1621 attaattttg gaactgtatg tgtgtgtcat acatcttcat agttacgagt ttaagatgga
     1681 tggaaatatc gatctaggat aggtatacat gttgatgtgg gttttactga tgcatataca
     1741 tgatggcata tgcagcatct attcatatgc tctaaccttg agtacctatc tattataata
     1801 aacaagtatg ttttataatt attttgatct tgatatactt ggatgatggc atatgcagca
     1861 gctatatgtg gattttttta gccctgcctt catacgctat ttatttgctt ggtactgttt
     1921 cttttgtcga tgctcaccct gttgtttggt gttacttctg cgccagtgcc aagttaattc
     1981 gagctcaagc tttaccgtgc gccacttgac acagactgga aatattctcg ttcacacagt
     2041 gctagggctt cagggtgtga gccgtggcgc atcttgtgga cttgccgtgg cgagctacct
     2101 ccacgctccc gcttccgcaa ttttgttgca gtgctttgga ttctcgtggt tgctcgtctc
     2161 aggaagtgaa gatgtcggcc tccaaggatg gcgaccctcg gatccagtat atctcggaca
     2221 gcatccgtac cattcctgat tttcctcaca aaggcattat gttccgagat gtgacgacgt
     2281 tgctgttgga tcataaggct ttcaaagaca cgatcgacat ctttgttgag cgttaccggg
     2341 accagaaggt ggacgtcatt gtgggaattg aagctcgagg tatttaaatt gattacgtac
     2401 agatgaacat ggcatcgtgg tgattgatga aactgctgct gtcggcttta acctctcttt
     2461 aggcattggt ttcgaagcgg gcaacaagcc gaaagaactg tacagcgaag aggcagtcaa
     2521 cggggaaact cagcaagcgc acttacaggc gattaaagag ctgatagcgc gtgacaaaaa
     2581 ccacccaagc gtggtgatgt ggagtattgc caacgaaccg gatacccgtc cgcaagtgca
     2641 cgggaatatt tcgccactgg cggaagcaac gcgtaaactc gacccgacgc gtccgatcac
     2701 ctgcgtcaat gtaatgttct gcgacgctca caccgatacc atcagcgatc tctttgatgc
     2761 ctaggtacgt aatcagttta aatacctcga gcttcaattc ccacaatgac gtccaccttc
     2821 tggtcccggt aacgctcaac aaagatgtcg atcgtgtctt tgaaagcctt atgatccaac
     2881 agcaacgtcg tcacatctcg gaacataatg cctttgtgag gaaaatcagg aatggtacgg
     2941 atgctgtccg agatatactg gatccgaggg tcgccatcct tggaggccga catcttcact
     3001 tcctgagacg agcaaccacg agaatccaaa gcactgcaac aaaattgcgg aagcgggagc
     3061 gtggaggtag ctcgccacgg caagtccaca agatgcgcca cggctcacac cctgaagccc
     3121 tagcactgtg tgaacgagaa tatttccagt ctgtgtcaag tggcgcacgg taaagcttgg
     3181 tacccctact ccaagatcgt tcaaacattt ggcaataaag tttcttaaga ttgaatcctg
     3241 ttgccggtct tgcgatgatt atcatataat ttctgttgaa ttacgttaag catgtaataa
     3301 ttaacatgta atgcatgacg ttatttatga gatgggtttt tatgattaga gtcccgcaat
     3361 tatacattta atacgcgata gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc
     3421 gcgcggtgtc atctatgtta ctagatcggg gatacagtct cagaagacca aagggctatt
     3481 gagacttttc aacaaagggt aatttcggga aacctcctcg gattccattg cccagctatc
     3541 tgtcacttca tcgaaaggac agtagaaaag gaaggtggct cctacaaatg ccatcattgc
     3601 gataaaggaa aggctatcat tcaagatgcc tctgccgaca gtggtcccaa agatggaccc
     3661 ccaccccgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
     3721 attgatgtga catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
     3781 acccttcctc tatataagga agttcatttc atttggagag gacagcccaa gcatggagaa
     3841 ccgacctgca ggcatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat
     3901 cgaaaagttc gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc
     3961 tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg
     4021 tttctacaaa gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga
     4081 agtgcttgac attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca
     4141 gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc
     4201 ggaggccatg gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt
     4261 cggaccgcaa ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga
     4321 tccccatgtg tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca
     4381 ggctctcgat gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca
     4441 cgcggatttc ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga
     4501 ctggagcgag gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag
     4561 gccgtggttg gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct
     4621 tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca
     4681 gagcttggtt gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat
     4741 cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt
     4801 ctggaccgat ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg
     4861 tccgagggca aaggaatgag cgggactctg gggttcggac ggtacgctga aatcaccagt
     4921 ctcttctaca aatctatctc tctctatttt ctccataaat aatgtgtgag tagtttcccg
     4981 ataagggaaa ttagggttct tatagggttt cgctcatgtg ttgagcatat aagaaaccct
     5041 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
     5101 aaatccagta ctaaaatcca gatctcctaa agtccctata gatctttgtc gtgaatataa
     5161 accagacacg agacgactaa acctggagcc cagacgccgt tcgaagctag aagtaccgct
     5221 taggcaggag gccgttaggg aaaagatgct aaggcagggt tggttacgtt gactcccccg
     5281 taggtttggt ttaaatatga tgaagtggac ggaaggaagg aggaagacaa ggaaggataa
     5341 ggttgcaggc cctgtgcaag gtaagaagat ggaaatttga tagaggtacg ctactatact
     5401 tatactatac gctaagggaa tgcttgtatt tataccctat accccctaat aaccccttat
     5461 caatttaaga aataatccgc ataagccccc gcttaaaaat tggtatcaga gccatgaata
     5521 ggtctatgac caaaactcaa gaggataaaa cctcaccaaa atacgaaaga gttcttaact
     5581 ctaaagataa aagatctttc aagatcaaaa ctagttccct cacaccggtg acggggatcg
     5641 catgcgatat ctgcgagaga tctagactgg cgtaatcatg gtcatagctg tttcctgtgt
     5701 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5761 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5821 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5881 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5941 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6001 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6061 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6121 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6181 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6241 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     6301 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6361 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6421 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6481 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     6541 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6601 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6661 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6721 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6781 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6841 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6901 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6961 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7021 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7081 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7141 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7201 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7261 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7321 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7381 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7441 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7501 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7561 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7621 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7681 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7741 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7801 ttccgcgcac atttccccga aaagtgc
//
