LOCUS       sequence-296579-        5201 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5201)
  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 5201)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pAGM8093
FEATURES             Location/Qualifiers
     source          1..5201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..1149
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    1295..1319
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    1464..1485
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1500..1505,1506..1523,1524..1530)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1538..1554
                     /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)."
     primer_bind     1543..1565
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1562..1578
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1562..1578
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1574..1897
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    1591..1647
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1648..1665)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1648..1664)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1657..1679)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1873..1892)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     misc_feature    2029..2053
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      2141..2851
                     /label=oriV
                     /note="incP origin of replication"
     CDS             3146..3937
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4278..4866
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4767..4786
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atgaatcgga cgtttgaccg gaaggcatac aggcaagaac tgatcgacgc ggggttttcc
       61 gccgaggatg ccgaaaccat cgcaagccgc accgtcatgc gtgcgccccg cgaaaccttc
      121 cagtccgtcg gctcgatggt ccagcaagct acggccaaga tcgagcgcga cagcgtgcaa
      181 ctggctcccc ctgccctgcc cgcgccatcg gccgccgtgg agcgttcgcg tcgtcttgaa
      241 caggaggcgg caggtttggc gaagtcgatg accatcgaca cgcgaggaac tatgacgacc
      301 aagaagcgaa aaaccgccgg cgaggacctg gcaaaacagg tcagcgaggc caagcaggcc
      361 gcgttgctga aacacacgaa gcagcagatc aaggaaatgc agctttcctt gttcgatatt
      421 gcgccgtggc cggacacgat gcgagcgatg ccaaacgaca cggcccgctc tgccctgttc
      481 accacgcgca acaagaaaat cccgcgcgag gcgctgcaaa acaaggtcat tttccacgtc
      541 aacaaggacg tgaagatcac ctacaccggc gtcgagctgc gggccgacga tgacgaactg
      601 gtgtggcagc aggtgttgga gtacgcgaag cgcaccccta tcggcgagcc gatcaccttc
      661 acgttctacg agctttgcca ggacctgggc tggtcgatca atggccggta ttacacgaag
      721 gccgaggaat gcctgtcgcg cctacaggcg acggcgatgg gcttcacgtc cgaccgcgtt
      781 gggcacctgg aatcggtgtc gctgctgcac cgcttccgcg tcctggaccg tggcaagaaa
      841 acgtcccgtt gccaggtcct gatcgacgag gaaatcgtcg tgctgtttgc tggcgaccac
      901 tacacgaaat tcatatggga gaagtaccgc aagctgtcgc cgacggcccg acggatgttc
      961 gactatttca gctcgcaccg ggagccgtac ccgctcaagc tggaaacctt ccgcctcatg
     1021 tgcggatcgg attccacccg cgtgaagaag tggcgcgagc aggtcggcga agcctgcgaa
     1081 gagttgcgag gcagcggcct ggtggaacac gcctgggtca atgatgacct ggtgcattgc
     1141 aaacgctagg gccttgtggg gtcagttccg gctgggggtt cagcagcccc tgctcggatc
     1201 tgttggaccg gacagtagtc atggttgatg ggctgcctgt atcgagtggt gattttgtgc
     1261 cgagctgccg gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca
     1321 aattgacgct tagacaactt aataacacat tgcggacgtt tttaatgtac tggggttgaa
     1381 cactctgtgg gtctcagagc ttgtcttcgc agctggcacg acaggtttcc cgactggaaa
     1441 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     1501 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     1561 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
     1621 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
     1681 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     1741 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     1801 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     1861 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     1921 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     1981 tgtgagaaga caagggacac gaagtgatcc gtttaaacta tcagtgtttg acaggatata
     2041 ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat ttaaaagggc
     2101 gtgaaaaggt ttatccgttc gtccatttgt atgtgccagc cgcctttgcg acgctcaccg
     2161 ggctggttgc cctcgccgct gggctggcgg ccgtctatgg ccctgcaaac gcgccagaaa
     2221 cgccgtcgaa gccgtgtgcg agacaccgcg gccgccggcg ttgtggatac ctcgcggaaa
     2281 acttggccct cactgacaga tgaggggcgg acgttgacac ttgaggggcc gactcacccg
     2341 gcgcggcgtt gacagatgag gggcaggctc gatttcggcc ggcgacgtgg agctggccag
     2401 cctcgcaaat cggcgaaaac gcctgatttt acgcgagttt cccacagatg atgtggacaa
     2461 gcctggggat aagtgccctg cggtattgac acttgagggg cgcgactact gacagatgag
     2521 gggcgcgatc cttgacactt gaggggcaga gtgctgacag atgaggggcg cacctattga
     2581 catttgaggg gctgtccaca ggcagaaaat ccagcatttg caagggtttc cgcccgtttt
     2641 tcggccaccg ctaacctgtc ttttaacctg cttttaaacc aatatttata aaccttgttt
     2701 ttaaccaggg ctgcgccctg tgcgcgtgac cgcgcacgcc gaaggggggt gccccccctt
     2761 ctcgaaccct cccggcccgc taacgcgggc ctcccatccc cccaggggct gcgcccctcg
     2821 gccgcgaacg gcctcacccc aaaaatggca gcgctggcca attcccgagg cacgaaccca
     2881 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
     2941 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
     3001 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     3061 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     3121 tcgccctaaa acaaagttaa acatcatggg ggaagcggtg atcgccgaag tatcgactca
     3181 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     3241 tttgtacggc tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt
     3301 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     3361 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     3421 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     3481 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     3541 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     3601 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     3661 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     3721 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     3781 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     3841 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta
     3901 cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctagctagaa attcgttcaa
     3961 gccgacgccg cttcgcggcg cggcttaact caagcgttag atgcactaag cacataattg
     4021 ctcacagcca aactatcagg tcaagtctgc ttttattatt tttaagcgtg cataataagc
     4081 cctacacaaa ttgggagata tatcatgctg tcagaccaag tttactcata tatactttag
     4141 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4201 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4261 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4321 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4381 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     4441 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4501 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4561 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4621 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4681 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4741 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4801 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4861 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg cagatcctag atgtggcgca
     4921 acgatgccgg cgacaagcag gagcgcaccg acttcttccg catcaagtgt tttggctctc
     4981 aggccgaggc ccacggcaag tatttgggca aggggtcgct ggtattcgtg cagggcaaga
     5041 ttcggaatac caagtacgag aaggacggcc agacggtcta cgggaccgac ttcattgccg
     5101 ataaggtgga ttatctggac accaaggcac caggcgggtc aaatcaggaa taagggcaca
     5161 ttgccccggc gtgagtcggg gcaatcccgc aaggagggtg a
//
