LOCUS       sequence-69040-e        5201 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 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 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: pAGM8067
FEATURES             Location/Qualifiers
     source          1..5201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    26..50
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      138..848
                     /label=oriV
                     /note="incP origin of replication"
     CDS             1143..1934
                     /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      2275..2863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2764..2783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3199..4347
                     /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    4493..4517
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    4662..4683
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4698..4703,4704..4721,4722..4728)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4736..4752
                     /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     4741..4763
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4760..4776
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4760..4776
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             4772..5095
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    4789..4845
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(4846..4863)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4846..4862)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4855..4877)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5071..5090)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtgatccgtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
       61 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
      121 catttgtatg tgccagccgc ctttgcgacg ctcaccgggc tggttgccct cgccgctggg
      181 ctggcggccg tctatggccc tgcaaacgcg ccagaaacgc cgtcgaagcc gtgtgcgaga
      241 caccgcggcc gccggcgttg tggatacctc gcggaaaact tggccctcac tgacagatga
      301 ggggcggacg ttgacacttg aggggccgac tcacccggcg cggcgttgac agatgagggg
      361 caggctcgat ttcggccggc gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc
      421 tgattttacg cgagtttccc acagatgatg tggacaagcc tggggataag tgccctgcgg
      481 tattgacact tgaggggcgc gactactgac agatgagggg cgcgatcctt gacacttgag
      541 gggcagagtg ctgacagatg aggggcgcac ctattgacat ttgaggggct gtccacaggc
      601 agaaaatcca gcatttgcaa gggtttccgc ccgtttttcg gccaccgcta acctgtcttt
      661 taacctgctt ttaaaccaat atttataaac cttgttttta accagggctg cgccctgtgc
      721 gcgtgaccgc gcacgccgaa ggggggtgcc cccccttctc gaaccctccc ggcccgctaa
      781 cgcgggcctc ccatcccccc aggggctgcg cccctcggcc gcgaacggcc tcaccccaaa
      841 aatggcagcg ctggccaatt cccgaggcac gaacccagtg gacataagcc tgttcggttc
      901 gtaagctgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
      961 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgggg
     1021 tacagtctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga
     1081 tgttatggag cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca
     1141 tcatggggga agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca
     1201 tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg
     1261 gcggcctgaa gccacacagc gatattgatt tgctggttac ggtgaccgta aggcttgatg
     1321 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
     1381 gcgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
     1441 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag
     1501 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
     1561 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
     1621 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
     1681 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
     1741 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
     1801 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
     1861 cctcgcgcgc agatcagttg gaagaatttg tccattacgt gaaaggcgag atcaccaagg
     1921 tagtcggcaa ataatgtcta gctagaaatt cgttcaagcc gacgccgctt cgcggcgcgg
     1981 cttaactcaa gcgttagatg cactaagcac ataattgctc acagccaaac tatcaggtca
     2041 agtctgcttt tattattttt aagcgtgcat aataagccct acacaaattg ggagatatat
     2101 catgctgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2161 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2221 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2281 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2341 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2401 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2461 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2521 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2581 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2641 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2701 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2761 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2821 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2881 ctttttacgg ttcctggcag atcctagatg tggcgcaacg atgccggcga caagcaggag
     2941 cgcaccgact tcttccgcat caagtgtttt ggctctcagg ccgaggccca cggcaagtat
     3001 ttgggcaagg ggtcgctggt attcgtgcag ggcaagattc ggaataccaa gtacgagaag
     3061 gacggccaga cggtctacgg gaccgacttc attgccgata aggtggatta tctggacacc
     3121 aaggcaccag gcgggtcaaa tcaggaataa gggcacattg ccccggcgtg agtcggggca
     3181 atcccgcaag gagggtgaat gaatcggacg tttgaccgga aggcatacag gcaagaactg
     3241 atcgacgcgg ggttttccgc cgaggatgcc gaaaccatcg caagccgcac cgtcatgcgt
     3301 gcgccccgcg aaaccttcca gtccgtcggc tcgatggtcc agcaagctac ggccaagatc
     3361 gagcgcgaca gcgtgcaact ggctccccct gccctgcccg cgccatcggc cgccgtggag
     3421 cgttcgcgtc gtcttgaaca ggaggcggca ggtttggcga agtcgatgac catcgacacg
     3481 cgaggaacta tgacgaccaa gaagcgaaaa accgccggcg aggacctggc aaaacaggtc
     3541 agcgaggcca agcaggccgc gttgctgaaa cacacgaagc agcagatcaa ggaaatgcag
     3601 ctttccttgt tcgatattgc gccgtggccg gacacgatgc gagcgatgcc aaacgacacg
     3661 gcccgctctg ccctgttcac cacgcgcaac aagaaaatcc cgcgcgaggc gctgcaaaac
     3721 aaggtcattt tccacgtcaa caaggacgtg aagatcacct acaccggcgt cgagctgcgg
     3781 gccgacgatg acgaactggt gtggcagcag gtgttggagt acgcgaagcg cacccctatc
     3841 ggcgagccga tcaccttcac gttctacgag ctttgccagg acctgggctg gtcgatcaat
     3901 ggccggtatt acacgaaggc cgaggaatgc ctgtcgcgcc tacaggcgac ggcgatgggc
     3961 ttcacgtccg accgcgttgg gcacctggaa tcggtgtcgc tgctgcaccg cttccgcgtc
     4021 ctggaccgtg gcaagaaaac gtcccgttgc caggtcctga tcgacgagga aatcgtcgtg
     4081 ctgtttgctg gcgaccacta cacgaaattc atatgggaga agtaccgcaa gctgtcgccg
     4141 acggcccgac ggatgttcga ctatttcagc tcgcaccggg agccgtaccc gctcaagctg
     4201 gaaaccttcc gcctcatgtg cggatcggat tccacccgcg tgaagaagtg gcgcgagcag
     4261 gtcggcgaag cctgcgaaga gttgcgaggc agcggcctgg tggaacacgc ctgggtcaat
     4321 gatgacctgg tgcattgcaa acgctagggc cttgtggggt cagttccggc tgggggttca
     4381 gcagcccctg ctcggatctg ttggaccgga cagtagtcat ggttgatggg ctgcctgtat
     4441 cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct ggtggcagga
     4501 tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa taacacattg cggacgtttt
     4561 taatgtactg gggttgaaca ctctgtgggt ctcattactt gtcttcgcag ctggcacgac
     4621 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     4681 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     4741 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc
     4801 ctgcaggtcg actctagagg atccccgggt accgagctcg aattcactgg ccgtcgtttt
     4861 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4921 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4981 gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     5041 tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     5101 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5161 atccgcttac agacaagctg tgagaagaca agggacacga a
//
