LOCUS       sequence-64259-e        3575 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 3575)
  TITLE     Murray Lab pBEST plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3575)
  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            "Unpublished"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-p15a-pBAD-TetO1-UTR1-deGFP-ssrA-T500
FEATURES             Location/Qualifiers
     source          1..3575
                     /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)"
     primer_bind     complement(66..87)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(327..346)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     674..695
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     rep_origin      1033..1577
                     /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     1478..1497
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1694..1711
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(1944..2735,2736..2804))
                     /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     2567..2586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2805..2909)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2977..2995
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     3472..3491
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    3515..3533
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3540..3558
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgga gcttttcact ggcgttgttc
       61 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      121 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      181 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
      241 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      301 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      361 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
      421 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
      481 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
      541 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      601 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
      661 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc gcagcaaacg
      721 acgaaaacta cgctttagct gcttaactcg agcaaagccc gccgaaaggc gggcttttct
      781 gtgtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
      841 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
      901 gccggcatga taagctgtca aacatgagaa ttacaactta tatcgtatgg ggctgacttc
      961 aggtgctaca tttgaagaga taaattgcac tgaaatctag aaatatttta tctgattaat
     1021 aagatgatct tcttgagatc gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa
     1081 aaccgccttg cagggcggtt tttcgaaggt tctctgagct accaactctt tgaaccgagg
     1141 taactggctt ggaggagcgc agtcaccaaa acttgtcctt tcagtttagc cttaaccggc
     1201 gcatgacttc aagactaact cctctaaatc aattaccagt ggctgctgcc agtggtgctt
     1261 ttgcatgtct ttccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1321 actgaacggg gggttcgtgc atacagtcca gcttggagcg aactgcctac ccggaactga
     1381 gtgtcaggcg tggaatgaga caaacgcggc cataacagcg gaatgacacc ggtaaaccga
     1441 aaggcaggaa caggagagcg cacgagggag ccgccagggg aaacgcctgg tatctttata
     1501 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     1561 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     1621 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     1681 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     1741 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     1801 tcatcagtgc caacatagta agccagtata cactccgcta gggtcatgag attatcaaaa
     1861 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     1921 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     1981 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2041 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2101 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2161 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2221 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2281 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2341 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2401 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2461 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     2521 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     2581 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     2641 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     2701 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     2761 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     2821 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     2881 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     2941 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3001 cgtcttcaag aattctggcg aatcctctga ccagccagaa aacgaccttt ctgtggtgaa
     3061 accggatgct gcaattcaga gcggcagcaa gtgggggaca gcagaagacc tgaccgccgc
     3121 agagtggatg tttgacatgg tgaagactat cgcaccatca gccagaaaac cgaattttgc
     3181 tgggtgggct aacgatatcc gcctgatgcg tgaacgtgac ggacgtaacc accgcgacat
     3241 gtgtgtgctg ttccgctggg catgcagaaa ccaattgtcc atattgcatc agacattgcc
     3301 gtcactgcgt cttttactgg ctcttctcgc taaccaaacc ggtaaccccg cttattaaaa
     3361 gcattctgta acaaagcggg accaaagcca tgacaaaaac gcgtaacaaa agtgtctata
     3421 atcacggcag aaaagtccac attgattatt tgcacggcgt cacactttgc tatgccatag
     3481 catttttatc cataagatta gcggatccta cctgtcccta tcagtgatag agattgacat
     3541 ccctatcagt gatagagata ctgagcacag ctagc
//
