LOCUS       sequence-64248-e        3723 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 3723)
  TITLE     Murray Lab pBEST plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3723)
  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            "Unpublished"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-PLtetO-1-UTR1-LacI-YbaQ-T500
FEATURES             Location/Qualifiers
     source          1..3723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             31..1110
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(50..69)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      1430..1974
                     /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     1875..1894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2091..2108
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(join(2341..3132,3133..3201))
                     /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     2964..2983
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3202..3306)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3374..3392
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    3663..3681
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3688..3706
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 aataattttg tttaacttta agaaggatcc atgaaaccag taacgttata cgatgtcgca
       61 gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt
      121 tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac
      181 cgcgtggcac aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt
      241 ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg
      301 ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg
      361 gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac
      421 caggatgcca ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc
      481 tctgaccaga cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc
      541 gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt
      601 tctgtctcgg cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt
      661 cagccgatag cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg
      721 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
      781 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
      841 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
      901 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
      961 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
     1021 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     1081 cgacaggttt cccgactgga aagcgggcag agaagggaag aaagagcaaa gaaggtagca
     1141 taactcgagc aaagcccgcc gaaaggcggg cttttctgtg tcgaccgatg cccttgagag
     1201 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     1261 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcatgataa gctgtcaaac
     1321 atgagaatta caacttatat cgtatggggc tgacttcagg tgctacattt gaagagataa
     1381 attgcactga aatctagaaa tattttatct gattaataag atgatcttct tgagatcgtt
     1441 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
     1501 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt
     1561 caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct
     1621 ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac
     1681 tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata
     1741 cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa
     1801 acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac
     1861 gagggagccg ccaggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac
     1921 tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc
     1981 tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc
     2041 cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag
     2101 tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct
     2161 tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc
     2221 cagtatacac tccgctaggg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2281 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2341 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     2401 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     2461 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     2521 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     2581 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     2641 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2701 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2761 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2821 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2881 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2941 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     3001 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     3061 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     3121 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     3181 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     3241 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     3301 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     3361 attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat tctggcgaat
     3421 cctctgacca gccagaaaac gacctttctg tggtgaaacc ggatgctgca attcagagcg
     3481 gcagcaagtg ggggacagca gaagacctga ccgccgcaga gtggatgttt gacatggtga
     3541 agactatcgc accatcagcc agaaaaccga attttgctgg gtgggctaac gatatccgcc
     3601 tgatgcgtga acgtgacgga cgtaaccacc gcgacatgtg tgtgctgttc cgctgggcat
     3661 gctccctatc agtgatagag attgacatcc ctatcagtga tagagatact gagcacagct
     3721 agc
//
