LOCUS       sequence-39586-e        6712 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6712)
  TITLE     Glazebrook/ Katagiri Lab Gateway destination vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6712)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pUNos_C1
FEATURES             Location/Qualifiers
     source          1..6712
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        13..2010
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     protein_bind    2053..2177
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2202..2207,2208..2225,2226..2232)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2286..2945
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2352..2371)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3287..3592
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     3471..3490
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(3633..3757)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      3823..4075
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(4093..4109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4093..4109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4106..4128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4117..4133
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(join(4141..4147,4148..4165,4166..4171))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4186..4207
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4324..4341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4495..5083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4575..4594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5254..6045,6046..6114))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5877..5896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6115..6219)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6287..6305
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6343..6365)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6465..6484
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6678..6700
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6692..6709
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6693..6709
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagcttgcat gcctgcagtg cagcgtgacc cggtcgtgcc cctctctaga gataatgagc
       61 attgcatgtc taagttataa aaaattacca catatttttt ttgtcacact tgtttgaagt
      121 gcagtttatc tatctttata catatattta aactttactc tacgaataat ataatctata
      181 gtactacaat aatatcagtg ttttagagaa tcatataaat gaacagttag acatggtcta
      241 aaggacaatt gagtattttg acaacaggac tctacagttt tatcttttta gtgtgcatgt
      301 gttctccttt ttttttgcaa atagcttcac ctatataata cttcatccat tttattagta
      361 catccattta gggtttaggg ttaatggttt ttatagacta atttttttag tacatctatt
      421 ttattctatt ttagcctcta aattaagaaa actaaaactc tattttagtt tttttattta
      481 ataatttaga tataaaatag aataaaataa agtgactaaa aattaaacaa atacccttta
      541 agaaattaaa aaaactaagg aaacattttt cttgtttcga gtagataatg ccagcctgtt
      601 aaacgccgtc gacgagtcta acggacacca accagcgaac cagcagcgtc gcgtcgggcc
      661 aagcgaagca gacggcacgg catctctgtc gctgcctctg gacccctctc gagagttccg
      721 ctccaccgtt ggacttgctc cgctgtcggc atccagaaat tgcgtggcgg agcggcagac
      781 gtgagccggc acggcaggcg gcctcctcct cctctcacgg caccggcagc tacgggggat
      841 tcctttccca ccgctccttc gctttccctt cctcgcccgc cgtaataaat agacaccccc
      901 tccacaccct ctttccccaa cctcgtgttg ttcggagcgc acacacacac aaccagatct
      961 cccccaaatc cacccgtcgg cacctccgct tcaaggtacg ccgctcgtcc tccccccccc
     1021 ccccctctct accttctcta gatcggcgtt ccggtccatg gttagggccc ggtagttcta
     1081 cttctgttca tgtttgtgtt agatccgtgt ttgtgttaga tccgtgctgc tagcgttcgt
     1141 acacggatgc gacctgtacg tcagacacgt tctgattgct aacttgccag tgtttctctt
     1201 tggggaatcc tgggatggct ctagccgttc cgcagacggg atcgatttca tgattttttt
     1261 tgtttcgttg catagggttt ggtttgccct tttcctttat ttcaatatat gccgtgcact
     1321 tgtttgtcgg gtcatctttt catgcttttt tttgtcttgg ttgtgatgat gtggtctggt
     1381 tgggcggtcg ttctagatcg gagtagaatt aattctgttt caaactacct ggtggattta
     1441 ttaattttgg atctgtatgt gtgtgccata catattcata gttacgaatt gaagatgatg
     1501 gatggaaata tcgatctagg ataggtatac atgttgatgc gggttttact gatgcatata
     1561 cagagatgct ttttgttcgc ttggttgtga tgatgtggtg tggttgggcg gtcgttcatt
     1621 cgttctagat cggagtagaa tactgtttca aactacctgg tgtatttatt aattttggaa
     1681 ctgtatgtgt gtgtcataca tcttcatagt tacgagttta agatggatgg aaatatcgat
     1741 ctaggatagg tatacatgtt gatgtgggtt ttactgatgc atatacatga tggcatatgc
     1801 agcatctatt catatgctct aaccttgagt acctatctat tataataaac aagtatgttt
     1861 tataattatt ttgatcttga tatacttgga tgatggcata tgcagcagct atatgtggat
     1921 ttttttagcc ctgccttcat acgctattta tttgcttggt actgtttctt ttgtcgatgc
     1981 tcaccctgtt gtttggtgtt acttctgcag gtcgactcta gaggatcccc tcgaggcgcg
     2041 ccaagctatc aaacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat
     2101 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     2161 catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt tatgcttccg
     2221 gctcgtataa tgtgtggatt ttgagttagg atccggcgag attttcagga gctaaggaag
     2281 ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta
     2341 aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc
     2401 tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct
     2461 ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg gcaatgaaag
     2521 acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa
     2581 ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca
     2641 tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta
     2701 ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa
     2761 acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa tattatacgc
     2821 aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct
     2881 tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg
     2941 cgtaatctag aggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     3001 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggtg
     3061 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     3121 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     3181 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     3241 attgaaatga acggctcttt tgctgacgag aacagggact ggtgaaatgc agtttaaggt
     3301 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     3361 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     3421 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     3481 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     3541 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     3601 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     3661 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     3721 tttacgtttc tcgttcagct ttcttgtaca aagtggttcg ataattcctt aattaactag
     3781 ttctagagcg gccgcccacc gcggtggagc tcgaatttcc ccgatcgttc aaacatttgg
     3841 caataaagtt tcttaagatt gaatcctgtt gccggtcttg cgatgattat catataattt
     3901 ctgttgaatt acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga
     3961 tgggttttta tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata
     4021 tagcgcgcaa actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgggaa
     4081 ttcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4141 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4201 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     4261 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     4321 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4381 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4441 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4501 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4561 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4621 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4681 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4741 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4801 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4861 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4921 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4981 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     5041 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     5101 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     5161 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5221 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5281 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5341 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     5401 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5461 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5521 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5581 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5641 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5701 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5761 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5821 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5881 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5941 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6001 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     6061 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     6121 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6181 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     6241 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6301 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     6361 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     6421 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     6481 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     6541 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     6601 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     6661 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg cc
//
