LOCUS       sequence-69048-e        3175 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 3175)
  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 3175)
  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: pICH50914
FEATURES             Location/Qualifiers
     source          1..3175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(509..526)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(509..525)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(518..540)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(734..753)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     853..875
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(913..931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        999..1103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1104..1172,1173..1964)
                     /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     complement(1322..1341)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2135..2723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2624..2643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2877..2894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3011..3032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3047..3052,3053..3070,3071..3077)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3085..3101
                     /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     3090..3112
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3109..3125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3109..3125
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tttgaagaca atgtgtgaga ccgaggatgc acatgtgacc gagggattgt cttcgtggaa
       61 acttgggact tcagattcca ccccttgcaa gcgtggaaag ccttccgacc aagggaagtg
      121 tcggatgtag aggaaatgga aagtttgttc tcagatgggg acctgcttga ttgcttcaca
      181 agaatgccag cttatgcggt aaacgcagag gaagatttag ctgcaatcag gaaaacgccc
      241 gagatggatg tcggtcaaga agttaaagag cctgcaggag acagaaatca atactcaaac
      301 cctgcagaaa ctttcctcaa caagctccac aggaaacaca gtagggaggt gaaacaccag
      361 gccgcaaaga aagctaaacg cctagctgaa atccaggagt caatgagagc tgaaggtgat
      421 gccgaaccaa atgaaataag cgggacgatg ggggcaatac ccagcaacgc cgaacttcct
      481 gggcccaaag ggtaccgagc tcgaattcac tggccgtcgt tttacaacgt cgtgactggg
      541 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
      601 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
      661 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat
      721 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
      781 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
      841 ctgtgaccat ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
      901 cgagtcgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
      961 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1021 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1081 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1141 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1201 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1261 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1321 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1381 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1441 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1501 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1561 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1621 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     1681 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1741 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     1801 ctggagccgg tgagcgtggt tctcgcggta tcattgcagc actggggcca gatggtaagc
     1861 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     1921 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     1981 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2041 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2281 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2881 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2941 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     3001 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     3061 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     3121 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcccc
//
