LOCUS       sequence-324555-        4747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose-inducible expression of green fluorescent protein encoding
            three amber codons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4747)
  AUTHORS   Ho JML, Miller CA, Parks SE, Mattia JR, Bennett MR
  TITLE     A suppressor tRNA-mediated feedforward loop eliminates leaky gene 
            expression in bacteria.
  JOURNAL   Nucleic Acids Res. 2021 Mar 18;49(5):e25. doi: 10.1093/nar/gkaa1179.
  PUBMED    33290521
REFERENCE   2  (bases 1 to 4747)
  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: "Nucleic 
            Acids Res."; date: "2021-03-18"; volume: "49(5):e25. doi"; pages: " 
            10.1093/nar/gkaa1179"
COMMENT     Sequence Label: pJH28_pBad_AraC_Para-sfGFP.3TAG
FEATURES             Location/Qualifiers
     source          1..4747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(569..1360,1361..1429))
                     /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     1192..1211
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1430..1521)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1540..1567)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1659..1745)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     1898..1915
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     CDS             complement(1992..2009)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(2767..3051)
                     /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     complement(2825..2844)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3078..3956
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(4112..4134)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4220..4360
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4275..4294
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4375..4392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(4557..4747,1..398))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4637..4656)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
       61 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
      121 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
      181 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
      241 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
      301 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      361 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
      421 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
      481 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
      541 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
      601 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
      661 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
      721 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
      781 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
      841 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
      901 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
      961 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1021 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     1081 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     1141 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg gataataccg
     1201 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     1261 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     1321 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     1381 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     1441 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     1501 gtatttagaa aaataaacaa aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg
     1561 gccttctgct taatttgatg cctggcagtt tatggcgggc gtcctgcccg ccaccctccg
     1621 ggccgttgct tcgcaacgtt caaatccgct cccggcggat ttgtcctact caggagagcg
     1681 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
     1741 atttgatgcc tggcagttcc ctactctcgc atggggagac cccacactac catcggcgct
     1801 acggcgtttc acttctgagt tcggcatggg gtcaggtggg accaccgcgc tactgccgcc
     1861 aggcaaattc tgttttatca gaccgcttct gcgttctgat ttaatctgta tcaggctgaa
     1921 aatcttctct catccgccaa aacagccaag cttcgaattc ccatatggta ccagctgcag
     1981 atctcgagtc aatgatgatg atgatgatgt gagcctttat acagttcgtc cataccgtgc
     2041 gtgatgccgg ccgcggtcac gaattccagc aggaccatgt ggtcacgctt ttcgttcgga
     2101 tctttactca gcacagactg cgtgctcagg taatgattgt ccggcagcag gaccgggcca
     2161 tcaccaatcg gggtgttttg ctgatagtgg tcggccagtt gaacgctgcc atcttccaca
     2221 ttatggcgga tcttgaagtt agccttgata ccgttcttct gcttatctgc ggtgatctac
     2281 acgttgtgcg aattgaagtt atattccagt ttatggccca gaatctaacc gtcttcctta
     2341 aaatcgatgc ctttcagttc aatacggtta accagcgtat caccttcgaa tttcacttcg
     2401 gcgcgcgttt tgtaggtgcc gtcatcttta aatgagatgg tacgttcttg gacataacct
     2461 tccggcatcg cggacttgaa aaagtcgtgg cgtttcatgt gatccgggta acgactgaaa
     2521 cactgaacac catacgtcag cgtggtgacc agggtcggcc acggaaccgg cagtttaccc
     2581 gtggtgcaaa taaacttcag cgtcagttta ccctaggttg catcgccttc accttcgccg
     2641 cgcacgctga atttgtgacc attgacatca ccatccagtt ccaccagaat cggcaccacg
     2701 cccgtaaaca gttcttcgcc cttgctcatc ctaggttaat tcctcctgtt agcccaaaaa
     2761 acgggtatgg agaaacagta gagagttgcg ataaaaagcg tcaggtagga tccgctaatc
     2821 ttatggataa aaatgctatg gcatagcaaa gtgtgacgcc gtgcaaataa tcaatgtgga
     2881 cttttctgcc gtgattatag acacttttgt tacgcgtttt tgtcatggct ttggtcccgc
     2941 tttgttacag aatgctttta ataagcgggg ttaccggttt ggttagcgag aagagccagt
     3001 aaaagacgca gtgacggcaa tgtctgatgc aatatggaca attggtttct tctctgaatg
     3061 gcgggagtat gaaaagtatg gctgaagcgc aaaatgatcc cctgctgccg ggatactcgt
     3121 ttaatgccca tctggtggcg ggtttaacgc cgattgaggc caacggttat ctcgattttt
     3181 ttatcgaccg accgctggga atgaaaggtt atattctcaa tctcaccatt cgcggtcagg
     3241 gggtggtgaa aaatcaggga cgagaatttg tttgccgacc gggtgatatt ttgctgttcc
     3301 cgccaggaga gattcatcac tacggtcgtc atccggaggc tcgcgaatgg tatcaccagt
     3361 gggtttactt tcgtccgcgc gcctactggc atgaatggct taactggccg tcaatatttg
     3421 ccaatacggg gttctttcgc ccggatgaag cgcaccagcc gcatttcagc gacctgtttg
     3481 ggcaaatcat taacgccggg caaggggaag ggcgctattc ggagctgctg gcgataaatc
     3541 tgcttgagca attgttactg cggcgcatgg aagcgattaa cgagtcgctc catccaccga
     3601 tggataatcg ggtacgcgag gcttgtcagt acatcagcga tcacctggca gacagcaatt
     3661 ttgatatcgc cagcgtcgca cagcatgttt gcttgtcgcc gtcgcgtctg tcacatcttt
     3721 tccgccagca gttagggatt agcgtcttaa gctggcgcga ggaccaacgt atcagccagg
     3781 cgaagctgct tttgagcacc acccggatgc ctatcgccac cgtcggtcgc aatgttggtt
     3841 ttgacgatca actctatttc tcgcgggtat ttaaaaaatg caccggggcc agcccgagcg
     3901 agttccgtgc cggttgtgaa gaaaaagtga atgatgtagc cgtcaagttg tcataattgg
     3961 taacgaatca gacaattgac ggcttgacgg agtagcatag ggtttgcaga atccctgctt
     4021 cgtccatttg acaggcacat tatgcatgcc gcttcgcctt cgcgcgcgaa ttgatctgct
     4081 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4141 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4201 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     4261 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     4321 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct
     4381 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4441 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4501 ccagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4561 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4621 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4681 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4741 gcgcttt
//
