LOCUS       Exported                6830 bp DNA     circular SYN 30-MAY-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6830)
  AUTHORS   Camilla Ciolli Mattioli
  TITLE     Direct Submission
  JOURNAL   Exported Jun 7, 2022 from SnapGene 6.0.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6830
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     981..1027
                     /label=202_intoAfeI_pBAD_FW
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1001..1027
                     /label=205_intoAfeI_pBAD_REV_flipped
     RBS             1312..1320
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             1329..2045
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1947..1982)
                     /label=461
     primer_bind     1965..1999
                     /label=460
     terminator      2364..2450
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2542..2569
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2563..2594)
                     /label=204_intoAfeI_pBAD_FW_flipped
     primer_bind     complement(2563..2587)
                     /label=203_intoAfeI_pBAD_REV
     promoter        2588..2679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2680..3540
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3582..4037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      4148..4736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5174..5219
                     /label=204_intoAfeI_pBAD_FW_flipped
     primer_bind     5174..5195
                     /label=202_intoAfeI_pBAD_FW
     primer_bind     5192..5219
                     /label=203_intoAfeI_pBAD_REV
     terminator      5213..5240
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5332..5418
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5649..6359)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     RBS             6368..6376
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(6403..6687)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(6661..6707)
                     /label=205_intoAfeI_pBAD_REV_flipped
     primer_bind     complement(6661..6690)
                     /label=202_intoAfeI_pBAD_FW
     primer_bind     complement(6687..6707)
                     /label=203_intoAfeI_pBAD_REV
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gtaaggaggt
     1321 tataaaaaat gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     1381 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     1441 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     1501 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
     1561 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     1621 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     1681 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     1741 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
     1801 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     1861 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     1921 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     1981 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     2041 aataaaggcg gacataaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaatgt ccagacctgc
     2101 aggcatgcaa gctttaatgc ggtagtttat cactagagtc gacctgcagg catgcaagct
     2161 tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     2221 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     2281 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     2341 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     2401 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     2461 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     2521 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     2581 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2641 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2701 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2761 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2821 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2881 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2941 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     3001 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     3061 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     3121 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     3181 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     3241 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     3301 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3361 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3421 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3481 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3541 ctgtcagacc aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc
     3601 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3661 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3721 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3781 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3841 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaacttg
     3901 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3961 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4021 attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4261 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     4321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4801 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4861 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4921 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtcat
     4981 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     5041 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     5101 accgtcatca ccgaaacgcg cgaggcagca aggagatggc gcccaacagt cccccggcca
     5161 cggggcctgc caccataccc acgccgaaac aagcagagtt tgtagaaacg caaaaaggcc
     5221 atccgtcagg atggccttct gcttaatttg atgcctggca gtttatggcg ggcgtcctgc
     5281 ccgccaccct ccgggccgtt gcttcgcaac gttcaaatcc gctcccggcg gatttgtcct
     5341 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
     5401 agcctttcgt tttatttgat gcctggcagt tccctactct cgcatgggga gaccccacac
     5461 taccatcggc gctacggcgt ttcacttctg agttcggcat ggggtcaggt gggaccaccg
     5521 cgctactgcc gccaggcaaa ttctgtttta tcagaccgct tctgcgttct gatttaatct
     5581 gtatcaggct gaaaatcttc tctcatccgc caaaacagcc aagcttgcat gcctgcaggt
     5641 cgactctatt acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
     5701 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg
     5761 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
     5821 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
     5881 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
     5941 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
     6001 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
     6061 cgctcccact tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
     6121 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
     6181 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
     6241 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
     6301 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatt
     6361 ttttataacc tccttacctc ctgctagccc aaaaaaacgg gtatggagaa acagtagaga
     6421 gttgcgataa aaagcgtcag gtaggatccg ctaatcttat ggataaaaat gctatggcat
     6481 agcaaagtgt gacgccgtgc aaataatcaa tgtggacttt tctgccgtga ttatagacac
     6541 ttttgttacg cgtttttgtc atggctttgg tcccgctttg ttacagaatg cttttaataa
     6601 gcggggttac cggtttggtt agcgagaaga gccagtaaaa gacgcagtga cggcaatgtc
     6661 tgatgcaata tggacaattg gtttcttgct catgagcccg aagtggcgag cccgatcttc
     6721 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     6781 ggccacgatg cgtccggcgt agaggatctg ctcatgtttg acagcttatc
//
