LOCUS       sequence-147242-        2198 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insulated MoClo promoter for E. coli. 36nt DNA spacer empirically 
            selected. Modified from BBa_R0040. [E:36INS-pTet:B].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2198)
  AUTHORS   Carr SB, Beal J, Densmore DM
  TITLE     Reducing DNA context dependence in bacterial promoters.
  JOURNAL   PLoS One. 2017 Apr 19;12(4):e0176013. doi: 
            10.1371/journal.pone.0176013. eCollection 2017.
  PUBMED    28422998
REFERENCE   2  (bases 1 to 2198)
  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: "PLoS One.";
            date: "2017-04-19"; volume: "12(4):e0176013. doi"; pages: " 
            10.1371/journal.pone.0176013. eCollection 2017"
COMMENT     Sequence Label: 36INS-pTet_EB
FEATURES             Location/Qualifiers
     source          1..2198
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    41..59
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    66..84
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    106..126
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     primer_bind     complement(132..149)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(303..891)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(383..402)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1062..1853,1854..1922))
                     /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     1685..1704
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1923..2027)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2095..2113
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2117..2160)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2163..2184
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gcttcggaga gaatacatag taaagaaacc tcactgttgt tccctatcag tgatagagat
       61 tgacatccct atcagtgata gagatactga gcactactag agacctacta gtagcggccg
      121 ctgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      181 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      241 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      301 tttttccaca ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      361 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      421 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      481 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      541 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      601 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      661 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      721 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      781 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      841 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      901 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
      961 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1021 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1081 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     1141 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     1201 cgcgacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     1261 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     1321 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     1381 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     1441 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     1501 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     1561 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     1621 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     1681 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     1741 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     1801 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     1861 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     1921 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     1981 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2041 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2101 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     2161 tggaattcgc ggccgcttct agagactagt gggtctca
//
