LOCUS       sequence-271922-        3240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the F1 Ampicillin resistance cassette as a type 8 part to be
            used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3240)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 3240)
  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: 
            "10.1021/acssynbio.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK8_004
FEATURES             Location/Qualifiers
     source          1..3240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     434..453
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(672..776)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      879..1467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1368..1387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1588..1606
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1613..1631
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1649..1660
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1667..2344
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2362..2433
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2449..2476
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      2506..2934
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2593..2612)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2803..2824
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3051..3240,1..602,603..671))
                     /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 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
       61 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      121 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      181 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      241 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      301 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      361 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      421 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      481 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      541 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      601 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      661 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      721 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      781 tttccccgaa aagtgccacc tgtcatgacc aaaatccctt aacgtgagtt ttcgttccac
      841 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
      901 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
      961 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1021 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1081 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1141 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1201 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1261 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1321 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1381 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1441 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     1501 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     1561 aaccgtgcgg ccgccccttg agaccagtcc ctatcagtga tagagattga catccctatc
     1621 agtgatagag atactgagca cggatctgaa agaggagaaa ggatctatgg cgagtagcga
     1681 agacgttatc aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca
     1741 cgagttcgaa atcgaaggtg aaggtgaagg tcgtccgtac gaaggtactc agaccgctaa
     1801 actgaaagtt accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt
     1861 ccagtacggt tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact
     1921 gtccttcccg gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt
     1981 taccgttacc caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg
     2041 tggtactaac ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc
     2101 ttccaccgaa cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct
     2161 gaaactgaaa gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa
     2221 accggttcag ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa
     2281 cgaagactac accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc
     2341 ttaataagga tctccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2401 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     2461 ggtgggcctt tctgcgttta taagtcggtc tcaccgagcg gccgcacgcg ccctgtagcg
     2521 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     2581 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     2641 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     2701 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     2761 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2821 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     2881 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2941 aaatattaac gcttacgatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3001 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3061 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3121 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3181 atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
//
