LOCUS       sequence-108431-        5861 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 5861)
  TITLE     Murray Lab pBEST plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5861)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBEST-OR2-OR1-Pr-araC, pBAD-TetR,
            pBAD-TetO1-d...
FEATURES             Location/Qualifiers
     source          1..5861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             87..965
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(1142..1159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1313..1901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1393..1412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2072..2863,2864..2932))
                     /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     2695..2714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2933..3037)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3123..3144)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3472..3491
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3731..3752
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             3789..3811
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3835..3853
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3860..3878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4442..4460
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             complement(4677..5300)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     5208..5227
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     RBS             5307..5329
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgca atatggacaa ttggtttctt
       61 ctctgaatgg cgggagtatg aaaagtatgg ctgaagcgca aaatgatccc ctgctgccgg
      121 gatactcgtt taatgcccat ctggtggcgg gtttaacgcc gattgaggcc aacggttatc
      181 tcgatttttt tatcgaccga ccgctgggaa tgaaaggtta tattctcaat ctcaccattc
      241 gcggtcaggg ggtggtgaaa aatcagggac gagaatttgt ttgccgaccg ggtgatattt
      301 tgctgttccc gccaggagag attcatcact acggtcgtca tccggaggct cgcgaatggt
      361 atcaccagtg ggtttacttt cgtccgcgcg cctactggca tgaatggctt aactggccgt
      421 caatatttgc caatacgggg ttctttcgcc cggatgaagc gcaccagccg catttcagcg
      481 acctgtttgg gcaaatcatt aacgccgggc aaggggaagg gcgctattcg gagctgctgg
      541 cgataaatct gcttgagcaa ttgttactgc ggcgcatgga agcgattaac gagtcgctcc
      601 atccaccgat ggataatcgg gtacgcgagg cttgtcagta catcagcgat cacctggcag
      661 acagcaattt tgatatcgcc agcgtcgcac agcatgtttg cttgtcgccg tcgcgtctgt
      721 cacatctttt ccgccagcag ttagggatta gcgtcttaag ctggcgcgag gaccaacgta
      781 tcagccaggc gaagctgctt ttgagcacca cccggatgcc tatcgccacc gtcggtcgca
      841 atgttggttt tgacgatcaa ctctatttct cgcgggtatt taaaaaatgc accggggcca
      901 gcccgagcga gttccgtgcc ggttgtgaag aaaaagtgaa tgatgtagcc gtcaagttgt
      961 cataactcga gcaaagcccg ccgaaaggcg ggcttttctg tgtcgaccga tgcccttgag
     1021 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     1081 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt
     1141 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1201 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1261 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1321 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtaagagg tggcgaaacc
     1381 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1441 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1501 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1561 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1621 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1681 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1741 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1801 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1861 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1921 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1981 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2041 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     2101 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2161 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2221 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2281 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2341 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2401 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2461 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2521 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2581 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2641 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     2701 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2761 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2821 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2881 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2941 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3001 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtccggg
     3061 ctttgctcga gttaagcagc taaagcgtag ttttcgtcgt ttgctgcgat cccggcggcg
     3121 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     3181 gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     3241 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg
     3301 aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg
     3361 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     3421 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     3481 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     3541 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag
     3601 gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     3661 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     3721 tggccgttta cgtcgccgtc cagctcgacc aggatgggaa caacgccagt gaaaagctcc
     3781 atggtatatc tccttcttaa agttaaacaa aattattgct agctgtgctc agtatctcta
     3841 tcactgatag ggatgtcaat ctctatcact gatagggaca ggtaggatcc gctaatctta
     3901 tggataaaaa tgctatggca tagcaaagtg tgacgccgtg caaataatca atgtggactt
     3961 ttctgccgtg attatagaca cttttgttac gcgtttttgt catggctttg gtcccgcttt
     4021 gttacagaat gcttttaata agcggggtta ccggtttggt tagcgagaag agccagtaaa
     4081 agacgcagtg acggcaatgt ctgatgcaat atggacaatt ggtttctgca tgcccagcgg
     4141 aacagcacac acatgtcgcg gtggttacgt ccgtcacgtt cacgcatcag gcggatatcg
     4201 ttagccccgg gatcgcagca aacgacgaaa actacgcttt agctgcttaa ctcgagcaaa
     4261 gcccgccgaa aggcgggctt ttctgtgtcg accgatgccc ttgagagcct tcaacccagt
     4321 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt
     4381 tatcatacta gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4441 aaataggcgt atcacgaggc cctttcgtct tcaagaattc tggcgaatcc tctgaccagc
     4501 cagaaaacga cctttctgtg gtgaaaccgg ggatcgcatg aaaaccgaat tttgctgggt
     4561 gggctaacga tatccgcatg atgcgtgaac gtgctatcgt cgccgcactt atgactgtct
     4621 tctttatcat tgacggcagt acagaaaagc ccgcctttcg gcgggctttg ctcgagttaa
     4681 gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc aaggccgaat
     4741 aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg taataatggc
     4801 ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg ctcttgatct
     4861 tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa tgcattctct
     4921 agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc atactgtttt
     4981 tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag taaagcacat
     5041 ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc taaagggcaa
     5101 aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa agcccgctta
     5161 ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc gagtttacgg
     5221 gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt aatcacttta
     5281 cttttatcta atctagacat ggtatatctc cttcttaaag ttaaacaaaa ttattgctag
     5341 ccaggtagga tccgctaatc ttatggataa aaatgctatg gcatagcaaa gtgtgacgcc
     5401 gtgcaaataa tcaatgtgga cttttctgcc gtgattatag acacttttgt tacgcgtttt
     5461 tgtcatggct ttggtcccgc tttgttacag aatgctttta ataagcgggg ttaccggttt
     5521 ggttagcgag aagagccagt aaaagacgca gtgacggcaa tgtctgatgc aatatggaca
     5581 attggtttct atgccgtaga atgctgcaat tcagagcgcc agcaagtggg ggacagcaga
     5641 agacctgacc gccgcagagt ggatgtttga catggtgaag actatcgcac catcagccag
     5701 aaaaccgaat tttgctgggt gggctaacga tatccgcctg atgcgtgaac gtgacggacg
     5761 taaccaccgc gacatgtgtg tgctgttccg ctgggcatgc tgagctaaca ccgtgcgtgt
     5821 tgacaatttt acctctggcg gtgataatgg ttgcagctag c
//
