LOCUS       sequence-65351-e        4527 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 4527)
  AUTHORS   Andreasson C, Schick AJ, Pfeiffer SM, Sarov M, Stewart F, Wurst W, 
            Schick JA
  TITLE     Direct cloning of isogenic murine DNA in yeast and relevance of 
            isogenicity for targeting in embryonic stem cells.
  JOURNAL   PLoS One. 2013 Sep 13;8(9):e74207. doi: 
            10.1371/journal.pone.0074207. eCollection 2013.
  PUBMED    24058528
REFERENCE   2  (bases 1 to 4527)
  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: "2013-09-13"; volume: "8(9):e74207. doi"; pages: " 
            10.1371/journal.pone.0074207. eCollection 2013"
COMMENT     Sequence Label: pRTVIR
FEATURES             Location/Qualifiers
     source          1..4527
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1094..1116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1105..1121
                     /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(1129..1135,1136..1153,1154..1159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(1312..1329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1483..2071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(2242..3033,3034..3102))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2865..2884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3103..3207)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3544..4526
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
       61 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
      121 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
      181 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
      241 cacatcacat ccgaacataa acaaccatga cagtcaacac taagacctat agtgagagag
      301 cagaaactca tgcctcacca gtagcacaac gattatttcg attaatggaa ctgaagaaaa
      361 ccaatttatg tgcatcaatt gatgttgata ccactaagga attccttgaa ttaattgata
      421 aattgggtcc ttatgtatgc ttaatcaaga ctcatattga tataatcaat gatttttcct
      481 atgaatccac tattgaacca ttattagaac tttcacgtaa acatcaattt atgatttttg
      541 aagatagaaa atttgctgat attggtaata ccgtgaagaa acaatatatt ggtggagttt
      601 ataaaattag tagttgggca gatattacta atgctcatgg tgtcactggg aatggagtag
      661 ttgaaggatt aaaacaggga gctaaagaaa ccaccaccaa ccaagagcca agagggttat
      721 tgatgttagc tgaattatca tcagtgggat cattagcata tggagaatat tctcaaaaaa
      781 ctgttgaaat tgctaaatcc gataaggaat ttgttattgg atttattgcc caacgtgata
      841 tgggtggaca agaagaagga tttgattggc ttattatgac acctggagtt ggattagatg
      901 ataaaggtga tggattagga caacaatata gaactgttga tgaagttgtt agcactggaa
      961 ctgatattat cattgttggt agaggattgt ttggtaaagg aagagatcca gatattgaag
     1021 gtaaaaggta tagagatgct ggttggaatg cttatttgta agcttctggc caattataag
     1081 atatcagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acataggagc
     1141 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg aggtaactca cattaattgc
     1201 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1261 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     1321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     1441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctcggccc
     1501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     1561 ataaagatac caggcgttcc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     1621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     1681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     1741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     1801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     1861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     1921 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     2101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     2161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     2221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     2281 ctgtctattt cgttcatcca tagttgcctg actgcccgtc gtgtagataa ctacgatacg
     2341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     2401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     2461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     2521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     2581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     2641 ccccatgttg tgaaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     2701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     2761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     2821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     2881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     2941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     3121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctggcttctc
     3241 aatgatattc gaatacgctt tgaggagata cagcctaata tccgacaaac tgttttacag
     3301 atttacgatc gtacttgtta cccatcattg aattttgaac atccgaacct gggagttttc
     3361 cctgaaacag atagtatatt tgaacctgta taataatata tagtctagcg ctttacggaa
     3421 gacaatgtat gtatttcggt tcctggagaa actattgcat ctattgcata ggtaatcttg
     3481 cacgtcgcat ccccggttca ttttctgcgt ttccatcttg cacttcaata gcatatcttt
     3541 gttaacgaag catctgtgct tcattttgta gaacaaaaat gcaacgcgag agcgctaatt
     3601 tttcaaacaa agaatctgag ctgcattttt acagaacaga aatgcaacgc gaaagcgcta
     3661 ttttaccaac gaagaatctg tgcttcattt ttgtaaaaca aaaatgcaac gcgagagcgc
     3721 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgagag
     3781 cgctatttta ccaacaaaga atctatactt cttttttgtt ctacaaaaat gcatcccgag
     3841 agcgctattt ttctaacaaa gcatcttaga ttactttttt tctcctttgt gcgctctata
     3901 atgcagtctc ttgataactt tttgcactgt aggtccgtta aggttagaag aaggctactt
     3961 tggtgtctat tttctcttcc ataaaaaaag cctgactcca cttcccgcgt ttactgatta
     4021 ctagcgaagc tgcgggtgca ttttttcaag ataaaggcat ccccgattat attctatacc
     4081 gatgtggatt gcgcatactt tgtgaacaga aagtgatagc gttgatgatt cttcattggt
     4141 cagaaaatta tgaacggttt cttctatttt gtctctatat actacgtata ggaaatgttt
     4201 acattttcgt attgttttcg attcactcta tgaatagttc ttactacaat ttttttgtct
     4261 aaagagtaat actagagata aacataaaaa atgtagaggt cgagtttaga tgcaagttca
     4321 aggagcgaaa ggtggatggg taggttatat agggatatag cacagagata tatagcaaag
     4381 agatactttt gagcaatgtt tgtggaagcg gtattcgcaa tattttagta gctcgttaca
     4441 gtccggtgcg tttttggttt tttgaaagtg cgtcttcaga gcgcttttgg ttttcaaaag
     4501 cgctctgaag ttcctatact ttctagc
//
