LOCUS       sequence-69066-e        4368 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 4368)
  AUTHORS   Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S
  TITLE     A modular cloning system for standardized assembly of multigene 
            constructs.
  JOURNAL   PLoS One. 2011 Feb 18;6(2):e16765. doi: 
            10.1371/journal.pone.0016765.
  PUBMED    21364738
REFERENCE   2  (bases 1 to 4368)
  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: "PLoS One.";
            date: "2011-02-18"; volume: "6(2):e16765. doi"; pages: " 
            10.1371/journal.pone.0016765"
COMMENT     Sequence Label: pICH54022
FEATURES             Location/Qualifiers
     source          1..4368
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    39..63
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(219..1367)
                     /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(1703..2291)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1783..1802)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2462..3253,3254..3322))
                     /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     3085..3104
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3323..3427)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3514..4145
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4237..4261
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 actattgtct tcacgaagtg gtttaaacta tcagtgtttg acaggatata ttggcgggta
       61 aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc gtgaaaaggt
      121 ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttccccaga tcaggcgctg
      181 gctgctgaac ccccagccgg aactgacccc acaaggccct agcgtttgca atgcaccagg
      241 tcatcattga cccaggcgtg ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg
      301 ccgacctgct cgcgccactt cttcacgcgg gtggaatccg atccgcacat gaggcggaag
      361 gtttccagct tgagcgggta cggctcccgg tgcgagctga aatagtcgaa catccgtcgg
      421 gccgtcggcg acagcttgcg gtacttctcc catatgaatt tcgtgtagtg gtcgccagca
      481 aacagcacga cgatttcctc gtcgatcagg acctggcaac gggacgtttt cttgccacgg
      541 tccaggacgc ggaagcggtg cagcagcgac accgattcca ggtgcccaac gcggtcggac
      601 gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac
      661 cggccattga tcgaccagcc caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc
      721 tcgccgatag gggtgcgctt cgcgtactcc aacacctgct gccacaccag ttcgtcatcg
      781 tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg
      841 accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag
      901 cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag
      961 gaaagctgca tttccttgat ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggcc
     1021 tcgctgacct gttttgccag gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt
     1081 cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg cctcctgttc aagacgacgc
     1141 gaacgctcca cggcggccga tggcgcgggc agggcagggg gagccagttg cacgctgtcg
     1201 cgctcgatct tggccgtagc ttgctggacc atcgagccga cggactggaa ggtttcgcgg
     1261 ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg
     1321 tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc gattcattca ccctccttgc
     1381 gggattgccc cgactcacgc cggggcaatg tgcccttatt cctgatttga cccgcctggt
     1441 gccttggtgt ccagataatc caccttatcg gcaatgaagt cggtcccgta gaccgtctgg
     1501 ccgtccttct cgtacttggt attccgaatc ttgccctgca cgaataccag cgaccccttg
     1561 cccaaatact tgccgtgggc ctcggcctga gagccaaaac acttgatgcg gaagaagtcg
     1621 gtgcgctcct gcttgtcgcc ggcatcgttg cgccacatct aggatctgcc aggaaccgta
     1681 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1741 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1801 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1861 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1921 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1981 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2041 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2101 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     2161 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2221 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2281 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2341 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2401 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2461 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2521 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2581 ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa
     2641 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2701 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2761 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2821 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2881 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2941 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3001 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3061 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3121 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3181 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3241 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3301 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3361 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3421 ttccgcgcac gaattggcca gcgctgccat ttttggggtg aggccgttcg cggccgaggg
     3481 gcgcagcccc tggggggatg ggaggcccgc gttagcgggc cgggagggtt cgagaagggg
     3541 gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc gcagccctgg ttaaaaacaa
     3601 ggtttataaa tattggttta aaagcaggtt aaaagacagg ttagcggtgg ccgaaaaacg
     3661 ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga cagcccctca aatgtcaata
     3721 ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt caaggatcgc gcccctcatc
     3781 tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc acttatcccc aggcttgtcc
     3841 acatcatctg tgggaaactc gcgtaaaatc aggcgttttc gccgatttgc gaggctggcc
     3901 agctccacgt cgccggccga aatcgagcct gcccctcatc tgtcaacgcc gcgccgggtg
     3961 agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc agtgagggcc aagttttccg
     4021 cgaggtatcc acaacgccgg cggccgcggt gtctcgcaca cggcttcgac ggcgtttctg
     4081 gcgcgtttgc agggccatag acggccgcca gcccagcggc gagggcaacc agcccggtga
     4141 gcgtcgcaaa ggagatcctg atctgactga tgggctgcct gtatcgagtg gtgattttgt
     4201 gccgagctgc cggtcgggga gctgttggct ggctggtggc aggatatatt gtggtgtaaa
     4261 caaattgacg cttagacaac ttaataacac attgcggacg tttttaatgt actggggtgg
     4321 atgcagtggg ccccactctg tgaagacaag caaggttttg acgggttt
//
