LOCUS       sequence-197799-        6706 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 6706)
  TITLE     Glazebrook/ Katagiri Lab Gateway destination vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6706)
  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            "Unpublished"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..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    988..1112
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1137..1142,1143..1160,1161..1167)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1221..1901
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(1287..1306)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2221..2526
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2405..2424
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2567..2691)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      2755..3007
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(3024..3040)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3024..3040)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3037..3059)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3048..3064
                     /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(3072..3078,3079..3096,3097..3102))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3117..3138
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3255..3272)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3426..4014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3506..3525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4185..4976,4977..5045))
                     /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     4808..4827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5046..5150)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5218..5236
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5274..5296)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5396..5415
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5609..5631
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5623..5640
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5624..5640
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(5656..6706,1..945)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
ORIGIN
        1 ggcccggtag ttctacttct gttcatgttt gtgttagatc cgtgtttgtg ttagatccgt
       61 gctgctagcg ttcgtacacg gatgcgacct gtacgtcaga cacgttctga ttgctaactt
      121 gccagtgttt ctctttgggg aatcctggga tggctctagc cgttccgcag acgggatcga
      181 tttcatgatt ttttttgttt cgttgcatag ggtttggttt gcccttttcc tttatttcaa
      241 tatatgccgt gcacttgttt gtcgggtcat cttttcatgc ttttttttgt cttggttgtg
      301 atgatgtggt ctggttgggc ggtcgttcta gatcggagta gaattaattc tgtttcaaac
      361 tacctggtgg atttattaat tttggatctg tatgtgtgtg ccatacatat tcatagttac
      421 gaattgaaga tgatggatgg aaatatcgat ctaggatagg tatacatgtt gatgcgggtt
      481 ttactgatgc atatacagag atgctttttg ttcgcttggt tgtgatgatg tggtgtggtt
      541 gggcggtcgt tcattcgttc tagatcggag tagaatactg tttcaaacta cctggtgtat
      601 ttattaattt tggaactgta tgtgtgtgtc atacatcttc atagttacga gtttaagatg
      661 gatggaaata tcgatctagg ataggtatac atgttgatgt gggttttact gatgcatata
      721 catgatggca tatgcagcat ctattcatat gctctaacct tgagtaccta tctattataa
      781 taaacaagta tgttttataa ttattttgat cttgatatac ttggatgatg gcatatgcag
      841 cagctatatg tggatttttt tagccctgcc ttcatacgct atttatttgc ttggtactgt
      901 ttcttttgtc gatgctcacc ctgttgtttg gtgttacttc tgcaggtcga ctctagagga
      961 tcccctcgag gcgcgccaag ctatcaaaca agtttgtaca aaaaagctga acgagaaacg
     1021 taaaatgata taaatatcaa tatattaaat tagattttgc ataaaaaaca gactacataa
     1081 tactgtaaaa cacaacatat ccagtcacta tggcggccgc attaggcacc ccaggcttta
     1141 cactttatgc ttccggctcg tataatgtgt ggattttgag ttaggatccg tcgagatttt
     1201 caggagctaa ggaagctaaa atggagaaaa aaatcactgg atataccacc gttgatatat
     1261 cccaatggca tcgtaaagaa cattttgagg catttcagtc agttgctcaa tgtacctata
     1321 accagaccgt tcagctggat attacggcct ttttaaagac cgtaaagaaa aataagcaca
     1381 agttttatcc ggcctttatt cacattcttg cccgcctgat gaatgctcat ccggaattcc
     1441 gtatggcaat gaaagacggt gagctggtga tatgggatag tgttcaccct tgttacaccg
     1501 ttttccatga gcaaactgaa acgttttcat cgctctggag tgaataccac gacgatttcc
     1561 ggcagtttct acacatatat tcgcaagatg tggcgtgtta cggtgaaaac ctggcctatt
     1621 tccctaaagg gtttattgag aatatgtttt tcgtctcagc caatccctgg gtgagtttca
     1681 ccagttttga tttaaacgtg gccaatatgg acaacttctt cgcccccgtt ttcaccatgg
     1741 gcaaatatta tacgcaaggc gacaaggtgc tgatgccgct ggcgattcag gttcatcatg
     1801 ccgtttgtga tggcttccat gtcggcagaa tgcttaatga attacaacag tactgcgatg
     1861 agtggcaggc ggggcgtaat ctagaggatc cggcttacta aaagccagat aacagtatgc
     1921 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
     1981 gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
     2041 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
     2101 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
     2161 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
     2221 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
     2281 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     2341 ctgctgtcag ataaagtccc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     2401 tggcgcatga tgaccaccga tatggccagt gtgccggtct ccgttatcgg ggaagaagtg
     2461 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     2521 atataaatgt caggctccct tatacacagc cagtctgcag gtcgaccata gtgactggat
     2581 atgttgtgtt ttacagtatt atgtagtctg ttttttatgc aaaatctaat ttaatatatt
     2641 gatatttata tcattttacg tttctcgttc agctttcttg tacaaagtgg ttcgataatt
     2701 cttaattaac tagttctaga gcggccgcca ccgcggtgga gctcgaattt ccccgatcgt
     2761 tcaaacattt ggcaataaag tttcttaaga ttgaatcctg ttgccggtct tgcgatgatt
     2821 atcatataat ttctgttgaa ttacgttaag catgtaataa ttaacatgta atgcatgacg
     2881 ttatttatga gatgggtttt tatgattaga gtcccgcaat tatacattta atacgcgata
     2941 gaaaacaaaa tatagcgcgc aaactaggat aaattatcgc gcgcggtgtc atctatgtta
     3001 ctagatcggg aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3061 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3121 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3181 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3241 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3301 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3361 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3421 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3481 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3541 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3601 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3661 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3721 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3781 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3841 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3901 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3961 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4021 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4081 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4141 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4201 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4261 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4321 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4381 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4441 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4501 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     4561 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4621 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4681 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4741 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4801 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4861 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4921 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4981 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5041 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5101 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5161 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     5221 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     5281 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     5341 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     5401 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     5461 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5521 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     5581 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     5641 gccaagcttg catgcctgca gtgcagcgtg acccggtcgt gcccctctct agagataatg
     5701 agcattgcat gtctaagtta taaaaaatta ccacatattt tttttgtcac acttgtttga
     5761 agtgcagttt atctatcttt atacatatat ttaaacttta ctctacgaat aatataatct
     5821 atagtactac aataatatca gtgttttaga gaatcatata aatgaacagt tagacatggt
     5881 ctaaaggaca attgagtatt ttgacaacag gactctacag ttttatcttt ttagtgtgca
     5941 tgtgttctcc tttttttttg caaatagctt cacctatata atacttcatc cattttatta
     6001 gtacatccat ttagggttta gggttaatgg tttttataga ctaatttttt tagtacatct
     6061 attttattct attttagcct ctaaattaag aaaactaaaa ctctatttta gtttttttat
     6121 ttaataattt agatataaaa tagaataaaa taaagtgact aaaaattaaa caaataccct
     6181 ttaagaaatt aaaaaaacta aggaaacatt tttcttgttt cgagtagata atgccagcct
     6241 gttaaacgcc gtcgacgagt ctaacggaca ccaaccagcg aaccagcagc gtcgcgtcgg
     6301 gccaagcgaa gcagacggca cggcatctct gtcgctgcct ctggacccct ctcgagagtt
     6361 ccgctccacc gttggacttg ctccgctgtc ggcatccaga aattgcgtgg cggagcggca
     6421 gacgtgagcc ggcacggcag gcggcctcct cctcctctca cggcaccggc agctacgggg
     6481 gattcctttc ccaccgctcc ttcgctttcc cttcctcgcc cgccgtaata aatagacacc
     6541 ccctccacac cctctttccc caacctcgtg ttgttcggag cgcacacaca cacaaccaga
     6601 tctcccccaa atccacccgt cggcacctcc gcttcaaggt acgccgctcg tcctcccccc
     6661 cccccctctc taccttctct agatcggcgt tccggtccat ggttag
//
