LOCUS       sequence-63381-e        4673 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 4673)
  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 4673)
  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: pBADmod1-linker-deGFP
FEATURES             Location/Qualifiers
     source          1..4673
                     /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"
     primer_bind     complement(349..370)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(610..629)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     957..978
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1003..1020
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1073..1090)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1243..1329
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1421..1448
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1468..1559
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1560..1628,1629..2420)
                     /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(1778..1797)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2591..3179
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3080..3099
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3333..3350
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3365..3505
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3431..3450)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3591..3613
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3769..4647)
                     /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 ggagcttttc actggcgttg ttcccatcct ggtcgagctg
      361 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      421 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      481 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      541 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      601 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      661 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      721 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
      781 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      841 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      901 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
      961 gtcctgctgg agttcgtgac cgccgccggg atctaactcg agcaccacca ccaccaccac
     1021 tgaattaagg tctccagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
     1081 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
     1141 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
     1201 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     1261 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     1321 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     1381 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
     1441 ggcctttttg cgtttctaca aactcttttt gtttattttt ctaaatacat tcaaatatgt
     1501 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     1561 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1621 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1681 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1741 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1801 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1861 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1921 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1981 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2041 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2101 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2161 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2221 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2281 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2341 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     2401 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     2461 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     2521 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     2581 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2641 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2701 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2761 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2821 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2881 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2941 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     3001 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     3061 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     3121 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     3181 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     3241 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     3301 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3361 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     3421 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     3481 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     3541 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     3601 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg
     3661 cgcgaaggcg aagcggcatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
     3721 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
     3781 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
     3841 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
     3901 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
     3961 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
     4021 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
     4081 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
     4141 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
     4201 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
     4261 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
     4321 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
     4381 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
     4441 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
     4501 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
     4561 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
     4621 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gag
//
