LOCUS       sequence-68983-e        2621 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 2621)
  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 2621)
  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: pAGM1311
FEATURES             Location/Qualifiers
     source          1..2621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    73..94
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(109..114,115..132,133..139)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    147..163
                     /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     152..174
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     171..187
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     171..187
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             183..506
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    200..256
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(257..274)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(257..273)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(266..288)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(482..501)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(639..657)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        725..829
                     /gene="bla"
                     /label=AmpR promoter
     CDS             830..1624
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      1795..2383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2284..2303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2537..2554
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtgaagacat agagaccgca gctggcacga caggtttccc gactggaaag cgggcagtga
       61 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
      121 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      181 ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagag gatccccggg
      241 taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
      301 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
      361 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga
      421 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca
      481 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
      541 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgacggtct
      601 cattgtcttc acgaagtggc tcttcagtgg acgaaagggc ctcgtgatac gcctattttt
      661 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
      721 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      781 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tggctaaaat
      841 gagaatatca ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga
      901 aggaatgtct cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa
      961 aatgacggac agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat
     1021 gctatggctg gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg
     1081 ctggagcaat ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga
     1141 tgaacaaagc cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc
     1201 catcgacata tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga
     1261 ttacttactg aataacgatc tggccgatgt ggattgcgaa aactgggaag aggacactcc
     1321 atttaaagat ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact
     1381 tgtcttttcc cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag
     1441 tggctttatt gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg
     1501 cgtccggtcg atcagggagg atatcgggga agaacagtat gtcgagctat tttttgactt
     1561 actggggatc aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt
     1621 ttagctgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     1681 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     1741 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     1801 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     1861 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     1921 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     1981 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2041 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2101 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2161 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2221 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2281 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2341 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2401 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2461 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     2521 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     2581 accgcctctc cccgcgcgtt ggccgattca ttaatcactc t
//
