LOCUS       sequence-254274-        6063 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses amcyan fluorescent gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  TITLE     A novel synthetic sRNA promoting protein overexpression in cell-free
            systems
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6063)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pART15-C1
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..722
                     /codon_start=1
                     /product="Anemonia majano cyan fluorescent protein"
                     /label=AmCyan
                     /translation="MALSNKFIGDDMKMTYHMDGCVNGHYFTVKGEGSGKPYEGTQTST
                     FKVTMANGGPLAFSFDILSTVFMYGNRCFTAYPTSMPDYFKQAFPDGMSYERTFTYEDG
                     GVATASWEISLKGNCFEHKSTFHGVNFPADGPVMAKMTTGWDPSFEKMTVCDGILKGDV
                     TAFLMLQGGGNYRCQFHTSYKTKKPVTMPPNHAVEHRIARTDLDKGGNSVQLTEHAVAH
                     ITSVVPF"
     primer_bind     complement(806..823)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      976..1062
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1154..1181
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    1281..1299
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(join(1306..1331,1332..1337,1338..1354,1355..1360,
                     1361..1379))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1336..1354
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1361..1379
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1515..1533
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1540..1558
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1950..2573
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(2023..2042)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     primer_bind     complement(2831..2848)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2965..3510)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3045..3064)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3675..4466,4467..4535))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4298..4317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4536..4627)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4675..4694
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4874..5752)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5779..6063
                     /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     5986..6005
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 actcgttttt ttgggctagc aggaggaaag ctatggctct ttcaaacaag tttatcggag
       61 atgacatgaa aatgacctac catatggatg gctgtgtcaa tgggcattac tttaccgtca
      121 aaggtgaagg cagcgggaag ccatacgaag ggacgcagac ctcgactttt aaagtcacca
      181 tggccaacgg tgggcccctt gcattctcct ttgacatact atctacagtg ttcatgtatg
      241 gaaatcgatg ctttactgcg tatcctacca gtatgcccga ctatttcaaa caagcatttc
      301 ctgacggaat gtcatatgaa aggactttta cctatgaaga tggaggagtt gctacagcca
      361 gttgggaaat aagccttaaa ggcaactgct ttgagcacaa atccacgttt catggagtga
      421 actttcctgc tgatggacct gtgatggcga agatgacaac tggttgggac ccatcttttg
      481 agaaaatgac tgtctgcgat ggaatattga agggtgatgt caccgcgttc ctcatgctgc
      541 aaggaggtgg caattacaga tgccaattcc acacttctta caagacaaaa aaaccggtga
      601 cgatgccacc aaaccatgcg gtggaacatc gcattgcgag gaccgacctt gacaaaggtg
      661 gcaacagtgt tcagcttacg gagcacgctg ttgcacatat aacctctgtt gtccctttct
      721 gacaccatgg tacccgggga tcctctagag tcgacctgca ggcatgcaag cttggctgtt
      781 ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc
      841 tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga
      901 actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag
      961 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     1021 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     1081 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     1141 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
     1201 ttgtttgggc gcctgtcact ttgcttgata tatgagaatt atttaacctt actcgagatc
     1261 tcctgctgat gtgctcagta tctctatcac tgatagggac agctgggtca gtgcgtcctg
     1321 ctgatgtgct cagtatctct atcactgata gggatgtcaa tctctatcac tgatagggac
     1381 agctgggatc ctaaatgaga attatttaac cttactcgag atctgatctt cagcatcttt
     1441 tactttcacc agccctgctg atgtgctcag aattatttaa ccttactcga gatctcctgc
     1501 tgatgtgctc agtatctcta tcactgatag ggatgtcaat ctctatcact gatagggaca
     1561 gctgggatcc taaatgagaa aaaagcaacg cactttaaat aagatacgtt gctttttcga
     1621 ttgatgaaca cctataatta aactattcat ctattattta tgattttttg tatatacaat
     1681 atttctagtt tgttaaagag aattaagaaa ataaatctcg aaaataataa agggaaaatc
     1741 agtttttgat atcaaaatta tacatgtcaa cgataataca aaatataata caaactataa
     1801 gatgttatca gtatttatta tgcatttaga ataaattttg tgtcgccctt tacacgtact
     1861 tagtcgctga acagctgatc aatgtggact tttctgccgg catgccagtt gtagatcagc
     1921 atagcataga caaccaagga ggaacagcta tgtctcgttt agataaaagt aaagtgatta
     1981 acagcgcatt agagctgctt aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg
     2041 cccagaagct aggtgtagag cagcctacat tgtattggca tgtaaaaaat aagcgggctt
     2101 tgctcgacgc cttagccatt gagatgttag ataggcacca tactcacttt tgccctttag
     2161 aaggggaaag ctggcaagat tttttacgta ataacgctaa aagttttaga tgtgctttac
     2221 taagtcatcg cgatggagca aaagtacatt taggtacacg gcctacagaa aaacagtatg
     2281 aaactctcga aaatcaatta gcctttttat gccaacaagg tttttcacta gagaatgcat
     2341 tatatgcact cagcgctgtg gggcatttta ctttaggttg cgtattggaa gatcaagagc
     2401 atcaagtcgc taaagaagaa agggaaacac ctactactga tagtatgccg ccattattac
     2461 gacaagctat cgaattattt gatcaccaag gtgcagagcc agccttctta ttcggccttg
     2521 aattgatcat ctgcggatta gaaaaacaac ttaaatgtga aagtgggtct taacagctgc
     2581 gtcaaaaggg cgacacccca taattagccc gggcgaaagg cccagtcttt cgactgagcc
     2641 tttcgtttta tttgatgcct gggagtcagc aacaccttct tcacgaggca gacctcagcg
     2701 ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt
     2761 catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat
     2821 attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa
     2881 tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag
     2941 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
     3001 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3061 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
     3121 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
     3181 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
     3241 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
     3301 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
     3361 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
     3421 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
     3481 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaaggg gtctgacgct
     3541 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3661 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3961 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
     4021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4261 cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc acatagcaga
     4321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4621 aaacaaatag gggttgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     4681 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     4741 tccggcgtag aggatctgct catgtttgac agcttatcat cgatgcataa tgtgcctgtc
     4801 aaatggacga agcagggatt ctgcaaaccc tatgctactc cgtcaagccg tcaattgtct
     4861 gattcgttac caattatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
     4921 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
     4981 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
     5041 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
     5101 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
     5161 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
     5221 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
     5281 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
     5341 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
     5401 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
     5461 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
     5521 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
     5581 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
     5641 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
     5701 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
     5761 tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
     5821 gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt aaaagcattc
     5881 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
     5941 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
     6001 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
     6061 cat
//
