LOCUS       sequence-236710-        3240 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes the F1 Ampicillin resistance cassette as a type 8a 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 Apr 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: MTK8a_004
FEATURES             Location/Qualifiers
     source          1..3240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      135..206
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      222..249
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      279..707
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(366..385)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     576..597
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(824..1615,1616..1684))
                     /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     1447..1466
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1685..1789)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1892..2480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2381..2400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2601..2619
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2626..2644
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2662..2673
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(2680..3240,1..117)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
ORIGIN
        1 cagctgccgg gtgcttacaa aaccgacatc aaactggaca tcacctccca caacgaagac
       61 tacaccatcg ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg tgcttaataa
      121 ggatctccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
      181 tctgttgttt gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc
      241 ctttctgcgt ttataagtcg gtctcaccga gcggccgcac gcgccctgta gcggcgcatt
      301 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
      361 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
      421 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
      481 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
      541 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
      601 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
      661 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
      721 aacgcttacg attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      781 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      841 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      901 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      961 cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1021 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1081 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1141 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1201 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1261 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1321 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1381 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1441 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1501 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1561 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1621 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1681 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1741 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1801 gaaaagtgcc acctgtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1861 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1921 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1981 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2041 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2101 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2161 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2221 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2281 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2341 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2401 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2461 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2521 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgtg
     2581 cggccgccaa ttgagaccag tccctatcag tgatagagat tgacatccct atcagtgata
     2641 gagatactga gcacggatct gaaagaggag aaaggatcta tggcgagtag cgaagacgtt
     2701 atcaaagagt tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg tcacgagttc
     2761 gaaatcgaag gtgaaggtga aggtcgtccg tacgaaggta ctcagaccgc taaactgaaa
     2821 gttaccaaag gtggtccgct gccgttcgct tgggacatcc tgtccccgca gttccagtac
     2881 ggttccaaag cttacgttaa acacccggct gacatcccgg actacctgaa actgtccttc
     2941 ccggaaggtt tcaaatggga acgtgttatg aacttcgaag acggtggtgt tgttaccgtt
     3001 acccaggact cctccctgca agacggtgag ttcatctaca aagttaaact gcgtggtact
     3061 aacttcccgt ccgacggtcc ggttatgcag aaaaaaacca tgggttggga agcttccacc
     3121 gaacgtatgt acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg tctgaaactg
     3181 aaagacggtg gtcactacga cgctgaagtt aaaaccacct acatggctaa aaaaccggtt
//
