LOCUS       sequence-69061-e        3173 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 3173)
  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 3173)
  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: pICH79289
FEATURES             Location/Qualifiers
     source          1..3173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..67)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(51..67)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(64..86)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    75..91
                     /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)."
     promoter        complement(join(99..105,106..123,124..129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    144..165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(282..299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(453..1041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(533..552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1212..2003,2004..2072))
                     /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     1835..1854
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2073..2177)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2245..2263
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2301..2323)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2423..2442
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2636..2658
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2650..2667
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2651..2667
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg
       61 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      421 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
      901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1201 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     1261 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     1321 ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc tccagattta
     1381 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     1441 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     1501 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     1561 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     1621 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     1681 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     1741 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     1801 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     1861 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     1921 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     1981 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     2041 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     2101 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     2161 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     2221 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg actcgcgcgt
     2281 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagatggt cacagcttgt
     2341 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2401 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     2461 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     2521 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     2581 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     2641 tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtaccctttg gtctcacaga
     2701 ttgtcttcga ggatgcacat gtgaccgagg gatgagaccg tggaaacttg ggacttcaga
     2761 ttccacccct tgcaagcgtg gaaagccttc cgaccaaggg aagtgtcgga tgtagaggaa
     2821 atggaaagtt tgttctcaga tggggacttg cttgattgct tcacaagaat gccagcttat
     2881 gcggtaaacg cagaggaaga tttagctgca atcaggaaaa cgcccgagat ggatgtcggt
     2941 caagaagtta aagaacctgc aggagacaga aatcaatact caaaccctgc agaaactttc
     3001 ctcaacaagc tccacaggaa acacagtagg gaggtgaaac accaggccgc aaagaaagct
     3061 aaacgcctag ctgaaatcca ggagtcaatg agagctgaag gtgatgccga accaaatgaa
     3121 ataagcgggg gcatgggggc aatacccagc aacgccgaac ttcctgggcc caa
//
