LOCUS       sequence-75064-e        2935 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 2935)
  AUTHORS   Engler C, Youles M, Gruetzner R, Ehnert TM, Werner S, Jones JD, 
            Patron NJ, Marillonnet S
  TITLE     A golden gate modular cloning toolbox for plants.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):839-43. doi: 10.1021/sb4001504. 
            Epub 2014 Feb 20.
  PUBMED    24933124
REFERENCE   2  (bases 1 to 2935)
  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"; doi: "10.1021/sb4001504";
            journalName: "ACS Synth Biol"; date: "2014-11-21- 21"; volume: "3"; 
            issue: "11"; pages: "839-43"
COMMENT     Sequence Label: pICH37326
FEATURES             Location/Qualifiers
     source          1..2935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(119..136)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(119..135)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(128..150)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(344..363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     463..485
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(523..541)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             933..1724
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      1895..2483
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2384..2403
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2637..2654
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2771..2792
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2807..2812,2813..2830,2831..2837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2845..2861
                     /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     2850..2872
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2869..2885
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2869..2885
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tttggtctca aatggctact caacgaaggg caaaccctag ctctctccat ctaattactg
       61 tattctctct gctcgtcgct gtcgtctcag gttgagaccg ggtaccgagc tcgaattcac
      121 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
      181 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
      241 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
      301 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
      361 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
      421 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
      481 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
      541 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgcgct
      721 cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt
      781 tcatggcttg ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc
      841 aagcgcgtta cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc
      901 agggcagtcg ccctaaaaca aagttaaaca tcatgaggga agcggtgatc gccgaagtat
      961 cgactcaact atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg
     1021 ccgtacattt gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt
     1081 tgctggttac ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc
     1141 ttttggaaac ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca
     1201 ttgttgtgca cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg
     1261 gagaatggca gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg
     1321 atctggctat cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg
     1381 cggaggaact ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct
     1441 taacgctatg gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt
     1501 tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg
     1561 actgggcaat ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg
     1621 cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg
     1681 tccactacgt gaaaggcgag atcaccaagg tagtcggcaa ataactgtca gaccaagttt
     1741 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     1801 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     1861 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1921 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1981 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2041 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2101 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2161 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2221 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2281 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2341 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2401 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2461 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2521 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2581 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2641 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2701 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     2761 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     2821 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     2881 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcccc
//
