LOCUS       sequence-462338-        3213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lvl 1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3213)
  AUTHORS   Duportet X, Wroblewska L, Guye P, Li Y, Eyquem J, Rieders J, 
            Rimchala T, Batt G, Weiss R
  TITLE     A platform for rapid prototyping of synthetic gene networks in 
            mammalian cells.
  JOURNAL   Nucleic Acids Res. 2014 Dec 1;42(21):13440-51. doi: 
            10.1093/nar/gku1082. Epub 2014 Nov 5.
  PUBMED    25378321
REFERENCE   2  (bases 1 to 3213)
  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"; doi: 
            "10.1093/nar/gku1082"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-1- 1"; volume: "42"; issue: "21"; pages: "13440-51"
COMMENT     Sequence Label: lvl1-S2S3 [backbone]
FEATURES             Location/Qualifiers
     source          1..3213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     606..625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(844..948)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1072..1115)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1118..1139
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    1140..1158
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1165..1183
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1202..1213
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1220..1897
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1942..2013
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2029..2056
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2097..2114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2437..3025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2517..2536)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3196..3213,1..774,775..843))
                     /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"
ORIGIN
        1 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
       61 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      121 gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      181 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      241 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      301 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      361 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      421 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      481 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      541 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      601 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      661 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      721 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      781 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      841 catactcttc ctttttcaat attattgaag catttatcaa ggttattgtc tcatgagcgg
      901 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca cgaattggcc
      961 agcgctgcat taccctgtta tccctaaccg gtgcgttttt atgcttgtag tattgtataa
     1021 tgtttttaag atccgctctt cacgcggagt gagaccgtat cacgaggcag aatttcagat
     1081 aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagagt
     1141 ccctatcagt gatagagatt gacatcccta tcagtgatag agatactgag cactactaga
     1201 gaaagaggag aaatactaga tggcttcctc cgaagacgtt atcaaagagt tcatgcgttt
     1261 caaagttcgt atggaaggtt ccgttaacgg tcacgagttc gaaatcgaag gtgaaggtga
     1321 aggtcgtccg tacgaaggta cccagaccgc taaactgaaa gttaccaaag gtggtccgct
     1381 gccgttcgct tgggacatcc tgtccccgca gttccagtac ggttccaaag cttacgttaa
     1441 acacccggct gacatcccgg actacctgaa actgtccttc ccggaaggtt tcaaatggga
     1501 acgtgttatg aacttcgaag acggtggtgt tgttaccgtt acccaggact cctccctgca
     1561 agacggtgag ttcatctaca aagttaaact gcgtggtacc aacttcccgt ccgacggtcc
     1621 ggttatgcag aaaaaaacca tgggttggga agcttccacc gaacgtatgt acccggaaga
     1681 cggtgctctg aaaggtgaaa tcaaaatgcg tctgaaactg aaagacggtg gtcactacga
     1741 cgctgaagtt aaaaccacct acatggctaa aaaaccggtt cagctgccgg gtgcttacaa
     1801 aaccgacatc aaactggaca tcacctccca caacgaagac tacaccatcg ttgaacagta
     1861 cgaacgtgct gaaggtcgtc actccaccgg tgcttaataa cgctgatagt gctagtgtag
     1921 atcgctacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     1981 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     2041 ggtgggcctt tctgcgttta tatactagta ggatcctaac tcgacgtgca ggcttcctcg
     2101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2161 gcggtaatgg tctcacgctt actgaagagc ggtctaatac ccaatctctc gtcttatcca
     2221 gatgttttat acgccttaat taatagggat aacagggtaa tatcttgccc tgcacgaata
     2281 ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga
     2341 tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc
     2401 tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2461 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2521 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2581 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2641 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2701 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2761 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2821 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2881 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2941 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3001 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3061 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3121 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3181 aacttggtct gacagttacc aatgcttaat cag
//
