LOCUS       sequence-462337-        3191 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 3191)
  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 3191)
  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-S4S5 [backbone]
FEATURES             Location/Qualifiers
     source          1..3191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(208..251)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    254..275
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    276..294
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    301..319
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             338..349
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             356..1033
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1078..1149
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1165..1192
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1233..1250)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1562..2150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1642..1661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2321..3112,3113..3181))
                     /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     2944..2963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(3182..3191,1..95))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcaaggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtt ccgcgcacga attggccagc gctgcattac
      121 cctgttatcc ctaaccggtg aattccctta tgtgagtgta aaaggcaggc gagtttgtcc
      181 cggagtgaga ccgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta
      241 aggatgattt ctggaattcg cggccgcttc tagagtccct atcagtgata gagattgaca
      301 tccctatcag tgatagagat actgagcact actagagaaa gaggagaaat actagatggc
      361 ttcctccgaa gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt
      421 taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca
      481 gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc
      541 cccgcagttc cagtacggtt ccaaagctta cgttaaacac ccggctgaca tcccggacta
      601 cctgaaactg tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaagacgg
      661 tggtgttgtt accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt
      721 taaactgcgt ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaaccatggg
      781 ttgggaagct tccaccgaac gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa
      841 aatgcgtctg aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat
      901 ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac
      961 ctcccacaac gaagactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc
     1021 caccggtgct taataacgct gatagtgcta gtgtagatcg ctactagagc caggcatcaa
     1081 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1141 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatata
     1201 ctagtaggat cctaactcga cgtgcaggct tcctcgctca ctgactcgct gcgctcggtc
     1261 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatggtctc acgctggttg
     1321 cttgcaaaag cagtaattgg aaagcactct caaagaatcc ttaattaata gggataacag
     1381 ggtaatatct tgccctgcac gaataccagc gaccccttgc ccaaatactt gccgtgggcc
     1441 tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg
     1501 gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     1981 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2461 gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     2941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3061 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3181 tactcttcct t
//
