LOCUS       Exported                8070 bp ds-DNA     circular SYN 17-OCT-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8070)
  AUTHORS   giselle
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Oct 24, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8070
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..120
                     /label=PBfP1E6
     regulatory      125..206
                     /regulatory_class="ribosome_binding_site"
                     /label=RBS1p-LP
     CDS             207..896
                     /codon_start=1
                     /label=mScarlet-I3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYWKQSFPEGFKWERVMIFEDGGT
                     VSVTQDTSLEDGTLIYKVKLRGGNFPPDGPVMQKRTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     terminator      919..946
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        989..1473
                     /label=PBT0779
     CDS             1474..2097
                     /codon_start=1
                     /label=TetR
                     /note="TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     terminator      2118..2146
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    2177..2284
                     /label=pTetR56
     protein_bind    2262..2280
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2285..2366
                     /label=RBS1P-LP
     CDS             2367..3086
                     /codon_start=1
                     /product="monomeric GFP derivative that displays 
                     exceptionally high brightness in cells (Campbell et al., 
                     2020)"
                     /label=mGreenLantern
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLGYGVACFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNSHKVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLK
                     ERVTAAGITHDMDELYK"
     terminator      3107..3134
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3384..4667)
                     /codon_start=1
                     /transl_table=11
                     /product="NBU integrase"
                     /label=NBU integrase
                     /protein_id="ARI71345.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKNGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTHKYALLSTHAGKRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     CDS             complement(5113..5973)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71346.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             6461..7195
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71348.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGNIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     rep_origin      7265..7653
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7824..7933
                     /note="incP origin of transfer"
ORIGIN
        1 caattgggct accttttttt tgttttgttt gcaatggtta atctattgtt aaaatttaaa
       61 gtttcacttg aactttcaaa taatgttctt atatttgcag tgtcgaaaga aacaaagtag
      121 gactgatcac acaacattta aaaaataaca ttatgaaagc actcgaaaag agattcaaag
      181 gtaatattaa caataattta ttttcaatgg atagcaccga ggcagtgatc aaggagttca
      241 tgcggttcaa ggtgcacatg gagggctcca tgaacggcca cgagttcgag atcgagggcg
      301 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
      361 gccccctgcc cttctcctgg gacatcctgt cccctcagtt catgtacggc tccagggcct
      421 tcatcaagca ccccgccgac atccccgact actggaagca gtccttcccc gagggcttca
      481 agtgggagcg cgtgatgatc ttcgaggacg gcggcaccgt gtccgtgacc caggacacct
      541 ccctggagga cggcaccctg atctacaagg tgaagctccg cggcggcaac ttccctcctg
      601 acggccccgt aatgcagaag aggacaatgg gctgggaagc atccaccgag cggttgtacc
      661 ccgaggacgt cgtgctgaag ggcgacatta agatggccct gcgcctgaag gacggcggcc
      721 gctacctggc ggacttcaag accacctaca aggccaagaa gcccgtgcag atgcccggcg
      781 ccttcaacat cgaccgcaag ttggacatca cctcccacaa cgaggactac accgtggtgg
      841 aacagtacga acgctccgtg gcccgccact ccaccggcgg ctccggtggc tcctaataaa
      901 tcctagctga ttagctcgag aaggccatcc tgacggatgg cctttttttt gtatggggac
      961 tctgagaacc ctacctttca aaccccgagg cagcatccat tgcctgaaaa ataaccagaa
     1021 ccgagtagcg gtcttgtgca taaagcatac tttgcagttc tgacagcttt tcgatatttt
     1081 gggctaactg ttctttggca tcctgtttgg acatatcagc agtaaatgag ggattgaaat
     1141 cggatacatg atgttccttc cccggtttgg ctgtaagctc ttttaagact tcttttttca
     1201 ttgttgtata attaggttgt tataggtata taacagttga ataaagaaat cagttcttgc
     1261 caaattaaga gcgttgaata gtaattcccc aattatgtat caacagattc tacataattc
     1321 cacactaatt ccacaattat atttatattg tctgctgttt atatgtttga ttatcagtta
     1381 tatatggcgg agtgaaacgt ttggcatctg ctttgctata tattaagtac aagaaacaat
     1441 agttctatta ttcacactta aaaaattacg attatgtcca gattagataa aagtaaagtg
     1501 attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac aacccgtaaa
     1561 ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa aaataagcgg
     1621 gctttgctcg acgccttagc cattgagatg ttagataggc accatactca cttttgccct
     1681 ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt tagatgtgct
     1741 ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac agaaaaacag
     1801 tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc actagagaat
     1861 gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt ggaagatcaa
     1921 gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat gccgccatta
     1981 ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt cttattcggc
     2041 cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg gtcttaatgg
     2101 ccacatcggt tcctgctaga aggcccatcc tgacggatgg ccttttggta ttcgagcatt
     2161 acctcatact ttggattttt gttttgtttg caatggttaa tctattgtta aaatttaaag
     2221 tttcacttga actttcaaat aatgttctta tatttgcagt gtccctatca gtgatagaga
     2281 gtaggatcac acaacattta aaaaataaca ttatgaaagc actcgaaaag agattcaaag
     2341 gtaatattaa caataattta ttttcaatgg tgagcaaggg cgaggagctg ttcaccgggg
     2401 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtccgcg
     2461 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     2521 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt aggctacggc gtggcctgct
     2581 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2641 gctacgtcca ggagcgcacc atctctttca aggacgacgg tacctacaag acccgcgccg
     2701 aggtgaagtt cgagggcgac accctggtga accgcatcgt gctgaagggc atcgacttca
     2761 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaaggtct
     2821 atatcacggc cgacaagcag aagaacggca tcaaggctaa cttcaagacc cgccacaacg
     2881 ttgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2941 gccccgtgct gctgcccgac aaccactacc tgagccatca gtccaagctg agcaaagacc
     3001 ccaacgagaa gcgcgatcac atggtcctga aggagagggt gaccgccgcc gggattacac
     3061 atgacatgga cgagctgtac aagtaatagc gttcagctcg actgtgagaa ggccatcctg
     3121 acggatggcc ttttttttcg ttataactag ttgcgttcct agccgctact agtgtaattg
     3181 cctatcttcc agtgatggaa cagcatttgt gcattggctg caacaatcag ccttacttgt
     3241 gcctgttcta tttccgaacc gaccgcttgt atgaatgcat caaaattcgt tttctctacg
     3301 ttggattcct tgttgctcat attgtgatga taatttctac aaatatagtc attggtaact
     3361 atctatgaaa ctgtttgata cttttatagt tgattaaact tgttcatggc atttgcctta
     3421 atatcatccg ctatgtcaat gtagggtttc atagctttgt agtcgctgtg tcccgtccat
     3481 ttcatgacca cctgtgccgg gattccgaga gccagcgcat tgcagatgaa tgtccttttt
     3541 cctgcatggg tactgagcaa agcgtatttg tgtgtgactt catcaatacg ttcatttccc
     3601 ttgtagtagg tttcccgtac aggctcgttg atttctgcca gttcgcccag ctctttcagg
     3661 taatcgttca tcttctggtt gctgatgacg ggcagagcca tgtaattctc gaaatggatg
     3721 tccttgtatt tgtccagtat ggctttgctg tatttgttca gttcaatcgt caggctgtcg
     3781 gcagtcttga ctgtggttat ttcgatgtgg tcggacttca catcgcttct tttcagattg
     3841 cgaacatccg aataccgcaa actcgtaaag cagcagaaca ggaaaacatc acgcacacgt
     3901 tccaggtatt gcttatcctt gggtatctgg tagtctttca gcttgttcag ttcatcccaa
     3961 gtcaggaaga ttactttttt cgaggtggtt ttcagtttcg gtttgaacgt atcgtatgca
     4021 atgttctgat gatgtccttt cttgaagctc cagcgcagga accatttgag gaatcccatt
     4081 tgcttgccga tggtgctgtt tctcatatcc ttggtgtcac gcaggaagtt gacgtattcg
     4141 ttcaatccaa actcgttgaa atagttgaac gttgcatcct ccttgaactc tttgaggtgg
     4201 ttcctcactg ctgcaaattt ttcataggtg gatgccgtcc agttattctg gttaccgcac
     4261 tcttttacaa actcatcgaa cacctcccaa aagctgacag gggcttcttc cggctgttct
     4321 tcactggtgt ctttcattct catgttgaaa gcttccttca actgttgggt cgttggcatg
     4381 acctcctgca cctcaaattc cttgaaaata ttctggattt cggcatagta tttcagcaag
     4441 tccgtattga tttcggctgc actttgcttt agcttgttgg tacatccgtt ctttacccgc
     4501 tgcttatctg catcccattt ggctacgtca atccggtagc ccgttgtaaa ctcgatacgt
     4561 tggctggcaa agatgacacg catacggatg ggtacgttct ctacgattgg cacaccgttc
     4621 tttttccggc tctccaatgc aaaaatgatg ttgcgcttga tattcataat tgggtgcgtt
     4681 tgaaattcta cacccaaata tacacccaat tattgagata gcaaaagaca tttagaaaca
     4741 tttactttta ctctatattg taatttacac ttgattatca gtcgtttgca gttttatgat
     4801 attctgtgaa agtataagtt cgagagcctg tctctccgca aaaaacgctg aaaatcagca
     4861 gattgcaaaa caaacaccct gttttacacc caagaatgta aagtcgggtg tttttgtttt
     4921 atttaagata atacaaccac tacataataa aagagtagcg atattaaaag aatccgatga
     4981 gaaaagacta atatttatct atccattcag tttgatttct caggacttta catcgtcctg
     5041 aaagtatttg ttggtaccgg taccgaggac gcgacagctg cagacgcgtt atgagtaaac
     5101 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5161 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5221 accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt
     5281 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5341 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5401 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5461 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5521 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5581 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5641 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5701 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5761 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5821 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5881 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5941 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat catggccgcg
     6001 ggattaaaag tcggggattg gtgaacaaaa aggtgtttct ctctttaaga gaaatatcgt
     6061 tttgctaaac agttgatatt gaggtatcat tttatcgtaa aagacatttt tgctcaacaa
     6121 ttgcttgacg gaaatcaaca aattttagca ttttgtaaaa aagtcgctat ataatttggt
     6181 gaattggagt tattttcata tttttgcatc ccgaagagtt tctcttaaag agagaaacat
     6241 cttttgcata ccttttccga ccgaattttt atgtcgtaaa gaggggcttt gcagggggtg
     6301 gactcagaaa gatgagaata gatgactatt gtagttgaaa cacatagaaa gttgctgata
     6361 tacagaccga tacgcatatc gggatgaacc atgagtacgt tcttttctca aaaaacataa
     6421 atattcgaaa agagatgcaa taaattaagg agaggttata atgaacaaag taaatataaa
     6481 agatagtcaa aattttatta cttcaaaata tcacatagaa aaaataatga attgcataag
     6541 tttagatgaa aaagataaca tctttgaaat aggtgcaggg aaaggtcatt ttactgctgg
     6601 attggtaaag agatgtaatt ttgtaacggc gatagaaatt gattctaaat tatgtgaggt
     6661 aactcgtaat aagctcttaa attatcctaa ctatcaaata gtaaatgatg atatactgaa
     6721 atttacattt cctagccaca atccatataa aatatttggc aacatacctt acaacataag
     6781 cacaaatata attcgaaaaa ttgtttttga aagttcagcc acaataagtt atttaatagt
     6841 ggaatatggt tttgctaaaa tgttattaga tacaaacaga tcactagcat tgctgttaat
     6901 ggcagaggta gatatttcta tattagcaaa aattcctagg tattatttcc atccaaaacc
     6961 taaagtggat agcacattaa ttgtattaaa aagaaagcca gcaaaaatgg catttaaaga
     7021 gagaaaaaaa tatgaaactt ttgtaatgaa atgggttaac aaagagtacg aaaaactgtt
     7081 tacaaaaaat caatttaata aagctttaaa acatgcgaga atatatgata taaacaatat
     7141 tagtttcgaa caatttgtat cgctatttaa tagttataaa atatttaacg gctaaaaaca
     7201 ataggccaca tgcaactgta aatgtttacg cgggtaccga caccgcggtg gaggggaatt
     7261 cccatgtcag ccgttaagtg ttcctgtgtc actcaaaatt gctttgagag gctctaaggg
     7321 cttctcagtg cgttacatcc ctggcttgtt gtccacaacc gttaaacctt aaaagcttta
     7381 aaagccttat atattctttt ttttcttata aaacttaaaa ccttagaggc tatttaagtt
     7441 gctgatttat attaatttta ttgttcaaac atgagagctt agtacgtgaa acatgagagc
     7501 ttagtacgtt agccatgaga gcttagtacg ttagccatga gggtttagtt cgttaaacat
     7561 gagagcttag tacgttaaac atgagagctt agtacgtgaa acatgagagc ttagtacgta
     7621 ctatcaacag gttgaactgc tgatcttcag atcctctacg ccggacgcat cgtggccgga
     7681 tcaattccgt tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
     7741 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
     7801 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
     7861 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
     7921 cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc tgatgaaacc
     7981 aagccaacca ggaagggcag cccacctatc acggaattga tccccctcga attggaattc
     8041 tagcctggaa tagggaagca ccacatcacc
//
