LOCUS       sequence-63378-e        3697 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 3697)
  AUTHORS   Shin J, Noireaux V
  TITLE     Efficient cell-free expression with the endogenous E. Coli RNA 
            polymerase and sigma factor 70.
  JOURNAL   J Biol Eng. 2010 Jun 24;4:8.
  PUBMED    20576148
REFERENCE   2  (bases 1 to 3697)
  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: "J Biol 
            Eng."; date: "2010-06-24"; volume: "4"; pages: "8"
COMMENT     Sequence Label: pBEST-p15A-PLlacO1-lacI-T500
FEATURES             Location/Qualifiers
     source          1..3697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             31..1113
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(50..69)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      1403..1947
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1848..1867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2064..2081
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(2314..3105,3106..3174))
                     /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     2937..2956
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3175..3279)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3347..3365
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    3662..3678
                     /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)."
ORIGIN
        1 aataattttg tttaacttta agaaggatcc atgaaaccag taacgttata cgatgtcgca
       61 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
      121 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
      181 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
      241 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
      301 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
      361 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
      421 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
      481 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
      541 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
      601 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
      661 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
      721 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
      781 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
      841 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
      901 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
      961 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     1021 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1081 cgacaggttt cccgactgga aagcgggcag tgagagctcg agcaaagccc gccgaaaggc
     1141 gggcttttct gtgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     1201 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     1261 taggacaggt gccggcatga taagctgtca aacatgagaa ttacaactta tatcgtatgg
     1321 ggctgacttc aggtgctaca tttgaagaga taaattgcac tgaaatctag aaatatttta
     1381 tctgattaat aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg
     1441 aaaacgaaaa aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt
     1501 tgaaccgagg taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc
     1561 cttaaccggc gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc
     1621 agtggtgctt ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg
     1681 cagcggtcgg actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac
     1741 ccggaactga gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc
     1801 ggtaaaccga aaggcaggaa caggagagcg cacgagggag ccgccagggg aaacgcctgg
     1861 tatctttata gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc
     1921 ttgtcagggg ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt
     1981 taagtatctt cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc
     2041 gccgcagtcg aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt
     2101 atcacatatt ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc
     2161 actgacaccc tcatcagtgc caacatagta agccagtata cactccgcta gggtcatgag
     2221 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     2281 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     2341 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2401 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2461 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2521 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2581 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2641 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2701 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2761 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2821 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2881 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2941 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     3001 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     3061 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     3121 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     3181 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     3241 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     3301 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3361 gaggcccttt cgtcttcaag aattctggcg aatcctctga ccagccagaa aacgaccttt
     3421 ctgtggtgaa accggatgct gcaattcaga gcggcagcaa gtgggggaca gcagaagacc
     3481 tgaccgccgc agagtggatg tttgacatgg tgaagactat cgcaccatca gccagaaaac
     3541 cgaattttgc tgggtgggct aacgatatcc gcctgatgcg tgaacgtgac ggacgtaacc
     3601 accgcgacat gtgtgtgctg ttccgctggg catgcataaa tgtgagcgga taacattgac
     3661 attgtgagcg gataacaaga tactgagcac agctagc
//
