LOCUS       sequence-473970-        5269 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Assembled plasmid containing four inserts (from POC1343, POC1344, 
            POC1345 and POC1346) assembled in POC1355.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5269)
  AUTHORS   Flores-Fernandez CN, Lin D, Robins K, O'Callaghan CA
  TITLE     UniClo: scarless hierarchical DNA assembly without sequence 
            constraint.
  JOURNAL   Nucleic Acids Res. 2025 Jun 20;53(12):gkaf548. doi: 
            10.1093/nar/gkaf548.
  PUBMED    40548934
REFERENCE   2  (bases 1 to 5269)
  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: "Nucleic 
            Acids Res."; date: "2025-06-20"; volume: "53(12):gkaf548. doi"; 
            pages: " 10.1093/nar/gkaf548"
COMMENT     Sequence Label: POC1358
FEATURES             Location/Qualifiers
     source          1..5269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    975..999
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1155..2303)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(2639..3227)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2719..2738)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3398..4189,4190..4258))
                     /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     4021..4040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4259..4363)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4450..5081
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5173..5197
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 cactctgtga cttggtctca tgccttgtgt tgatactaat gcgttaatgt tcggggcaag
       61 gggttctacg tggctagtta aataggtgcg tgatgcaaga tttatactag atttttcaaa
      121 tttggtccgt atcttggaaa catggggctg gccgtccact atcttggtat gcggtcgttg
      181 tactgggaaa ttaagatgcc attgttggta ctcacgtgtg tatatgaggc tgtttgtatc
      241 catgtagtga cggtggagta acaagatgtt acgaagtagg aatttcgggg taggtatcga
      301 aggattagat cattcaggcc ttcttgcatc cggctcagcc tgtgtactag agcgtgggaa
      361 tgcaaatacg tccgtggttg cggatctggg ccgcggaccg gtgggtgcgc cttctggtgt
      421 tgggcggtct ttgaggcaga cagtgaggtc tagagttggt tgatctgtag ctaagacgtc
      481 acctcagagt ttatcagtac tgtagacttt tcattcaagg tacccctggt cgaaaatacg
      541 cgtggtagaa tcgttgttgc gcgtatcctt tttaagacgg aagcgcaccg gggggaatgg
      601 tgtggttaag agggttggca taacgctagg catggtggtt acgtactgta ttcgacccat
      661 tcggtgatgt catatggcgt ggtgcccaga cgacgctaga ccgtagacta tggtcattgt
      721 gggaaggaat cgcgacttgc ggttctcacg gttgagagtg gaccggccga gctgacattc
      781 taacgagtcg ttgccacttc tttgcttatt acttcataaa ctccaaattc ggcatgatcg
      841 gagtcggcga gcagtcagta ccggggcgat gtgggttaag gacgccaagt tttccatgca
      901 gaacacgtgg cggtcatgac gtcgatcggg tacagatgag accaagtcac gaagtggttt
      961 aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat
     1021 tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt
     1081 gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc agccggaact
     1141 gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca ggcgtgttcc
     1201 accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc
     1261 acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag cgggtacggc
     1321 tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac
     1381 ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg
     1441 atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc
     1501 agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt
     1561 aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga ccagcccagg
     1621 tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg
     1681 tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc gacgccggtg
     1741 taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc
     1801 gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct
     1861 cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc
     1921 tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc
     1981 tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac
     2041 ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc ggccgatggc
     2101 gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc
     2161 tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt
     2221 gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc
     2281 ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac tcacgccggg
     2341 gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag ataatccacc
     2401 ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta cttggtattc
     2461 cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc gtgggcctcg
     2521 gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt gtcgccggca
     2581 tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2641 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2701 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2761 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2821 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2881 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2941 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     3001 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     3061 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3121 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3181 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3241 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3301 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3361 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3421 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3481 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3541 aaccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3601 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3661 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3721 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3781 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3841 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3901 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3961 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4021 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4081 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4141 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4201 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4261 tcttcctttt tcaatattat tgaagcattt atcaaggtta ttgtctcatg agcggataca
     4321 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacgaat tggccagcgc
     4381 tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag
     4441 gcccgcgtta gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg
     4501 gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag
     4561 caggttaaaa gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg
     4621 attttctgcc tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac
     4681 tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt
     4741 gtcaataccg cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt
     4801 aaaatcaggc gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc
     4861 gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt
     4921 ccgcccctca tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc
     4981 cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg
     5041 ccgccagccc agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag atcctgatct
     5101 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
     5161 ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa
     5221 taacacattg cggacgtttt taatgtactg gggtggatgc agtgggccc
//
