LOCUS       sequence-70396-e        4292 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 4292)
  AUTHORS   Sun ZZ, Yeung E, Hayes CA, Noireaux V, Murray RM
  TITLE     Linear DNA for rapid prototyping of synthetic biological circuits in
            an Escherichia coli based TX-TL cell-free system.
  JOURNAL   ACS Synth Biol. 2014 Jun 20;3(6):387-97. doi: 10.1021/sb400131a. 
            Epub 2013 Dec 4.
  PUBMED    24303785
REFERENCE   2  (bases 1 to 4292)
  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"; doi: "10.1021/sb400131a";
            journalName: "ACS Synth Biol"; date: "2014-06-20- 20"; volume: "3"; 
            issue: "6"; pages: "387-97"
COMMENT     Sequence Label: pBADmod1-linker2-gamS
FEATURES             Location/Qualifiers
     source          1..4292
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             622..639
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(692..709)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      862..948
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1040..1067
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1087..1178
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1179..1247,1248..2039)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1397..1416)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2210..2798
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2699..2718
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2952..2969
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2984..3124
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3050..3069)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3210..3232
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3388..4266)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttaact
      301 ttaagaagga gatataccat gggaaacgct tattacattc aggatcgtct tgaggctcag
      361 agctgggcgc gtcactacca gcagctcgcc cgtgaagaga aagaggcaga actggcagac
      421 gacatggaaa aaggcctgcc ccagcacctg tttgaatcgc tatgcatcga tcatttgcaa
      481 cgccacgggg ccagcaaaaa atccattacc cgtgcgtttg atgacgatgt tgagtttcag
      541 gagcgcatgg cagaacacat ccggtacatg gttgaaacca ttgctcacca ccaggttgat
      601 attgattcag aggtactcga gcaccaccac caccaccact gaattaaggt ctccagcttg
      661 gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa
      721 gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca
      781 tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga
      841 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
      901 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
      961 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     1021 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     1081 actctttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1141 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1201 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1261 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1321 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1381 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     1441 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1501 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1561 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     1621 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     1681 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     1741 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     1801 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     1861 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     1921 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     1981 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2041 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2101 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2161 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2221 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2281 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2341 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2401 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2461 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2521 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2581 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2641 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2701 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2761 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2821 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2881 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     2941 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3001 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     3061 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3121 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3181 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3241 gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc
     3301 ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa
     3361 gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt
     3421 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     3481 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     3541 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     3601 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     3661 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     3721 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     3781 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     3841 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     3901 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     3961 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     4021 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     4081 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     4141 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     4201 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc
     4261 agccatactt ttcatactcc cgccattcag ag
//
