LOCUS       Exported                8059 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 8059)
  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..8059
                     /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..1462
                     /label=PBT2364
     CDS             1463..2086
                     /codon_start=1
                     /label=TetR
                     /note="TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     terminator      2107..2135
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    2166..2273
                     /label=pTetR56
     protein_bind    2251..2269
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2274..2355
                     /label=RBS1P-LP
     CDS             2356..3075
                     /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      3096..3123
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3373..4656)
                     /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(5102..5962)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71346.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             6450..7184
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71348.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGNIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     misc_feature    7250..8023
                     /label=Origin
     rep_origin      7254..7642
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7813..7922
                     /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 aaccccgagc ggcatgacgg ttcgtaacgt attagacgaa
     1021 agaggacgcg aactatgcgg tgaatattgc cgtttctacg atttgaaacg gacagggatg
     1081 ttcaagagtt ccgattatct gaaagatact cacccggatc tcgcacggtt ctttaatccg
     1141 aattatgctt tacgtcccat ttctacgacc tttaccgcta ccattatcaa cggcagtgag
     1201 tatcagaatc cgggatatta aattgtaaga cctatatacc caatcattat cggtacgata
     1261 atccatatag ccacgctatt tatcgtacca atgattgggg atattttcca cacctttcca
     1321 catttttcca cacctcccca ataatatcgt ttttttcaac tctgctgtta atctgtatct
     1381 tatcctttca gcatttcttt atatcctatt ggtatatcct ttgtatcgac tttatagcaa
     1441 atgtattttt aaatataaaa gtatgtccag attagataaa agtaaagtga ttaacagcgc
     1501 attagagctg cttaatgagg tcggaatcga aggtttaaca acccgtaaac tcgcccagaa
     1561 gctaggtgta gagcagccta cattgtattg gcatgtaaaa aataagcggg ctttgctcga
     1621 cgccttagcc attgagatgt tagataggca ccatactcac ttttgccctt tagaagggga
     1681 aagctggcaa gattttttac gtaataacgc taaaagtttt agatgtgctt tactaagtca
     1741 tcgcgatgga gcaaaagtac atttaggtac acggcctaca gaaaaacagt atgaaactct
     1801 cgaaaatcaa ttagcctttt tatgccaaca aggtttttca ctagagaatg cattatatgc
     1861 actcagcgct gtggggcatt ttactttagg ttgcgtattg gaagatcaag agcatcaagt
     1921 cgctaaagaa gaaagggaaa cacctactac tgatagtatg ccgccattat tacgacaagc
     1981 tatcgaatta tttgatcacc aaggtgcaga gccagccttc ttattcggcc ttgaattgat
     2041 catatgcgga ttagaaaaac aacttaaatg tgaaagtggg tcttaatggc cacatcggtt
     2101 cctgctagaa ggcccatcct gacggatggc cttttggtat tcgagcatta cctcatactt
     2161 tggatttttg ttttgtttgc aatggttaat ctattgttaa aatttaaagt ttcacttgaa
     2221 ctttcaaata atgttcttat atttgcagtg tccctatcag tgatagagag taggatcaca
     2281 caacatttaa aaaataacat tatgaaagca ctcgaaaaga gattcaaagg taatattaac
     2341 aataatttat tttcaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2401 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtccgcgg cgagggcgag
     2461 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2521 gtgccctggc ccaccctcgt gaccacctta ggctacggcg tggcctgctt cgcccgctac
     2581 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2641 gagcgcacca tctctttcaa ggacgacggt acctacaaga cccgcgccga ggtgaagttc
     2701 gagggcgaca ccctggtgaa ccgcatcgtg ctgaagggca tcgacttcaa ggaggacggc
     2761 aacatcctgg ggcacaagct ggagtacaac ttcaacagcc acaaggtcta tatcacggcc
     2821 gacaagcaga agaacggcat caaggctaac ttcaagaccc gccacaacgt tgaggacggc
     2881 ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     2941 ctgcccgaca accactacct gagccatcag tccaagctga gcaaagaccc caacgagaag
     3001 cgcgatcaca tggtcctgaa ggagagggtg accgccgccg ggattacaca tgacatggac
     3061 gagctgtaca agtaatagcg ttcagctcga ctgtgagaag gccatcctga cggatggcct
     3121 tttttttcgt tataactagt tgcgttccta gccgctacta gtgtaattgc ctatcttcca
     3181 gtgatggaac agcatttgtg cattggctgc aacaatcagc cttacttgtg cctgttctat
     3241 ttccgaaccg accgcttgta tgaatgcatc aaaattcgtt ttctctacgt tggattcctt
     3301 gttgctcata ttgtgatgat aatttctaca aatatagtca ttggtaacta tctatgaaac
     3361 tgtttgatac ttttatagtt gattaaactt gttcatggca tttgccttaa tatcatccgc
     3421 tatgtcaatg tagggtttca tagctttgta gtcgctgtgt cccgtccatt tcatgaccac
     3481 ctgtgccggg attccgagag ccagcgcatt gcagatgaat gtcctttttc ctgcatgggt
     3541 actgagcaaa gcgtatttgt gtgtgacttc atcaatacgt tcatttccct tgtagtaggt
     3601 ttcccgtaca ggctcgttga tttctgccag ttcgcccagc tctttcaggt aatcgttcat
     3661 cttctggttg ctgatgacgg gcagagccat gtaattctcg aaatggatgt ccttgtattt
     3721 gtccagtatg gctttgctgt atttgttcag ttcaatcgtc aggctgtcgg cagtcttgac
     3781 tgtggttatt tcgatgtggt cggacttcac atcgcttctt ttcagattgc gaacatccga
     3841 ataccgcaaa ctcgtaaagc agcagaacag gaaaacatca cgcacacgtt ccaggtattg
     3901 cttatccttg ggtatctggt agtctttcag cttgttcagt tcatcccaag tcaggaagat
     3961 tacttttttc gaggtggttt tcagtttcgg tttgaacgta tcgtatgcaa tgttctgatg
     4021 atgtcctttc ttgaagctcc agcgcaggaa ccatttgagg aatcccattt gcttgccgat
     4081 ggtgctgttt ctcatatcct tggtgtcacg caggaagttg acgtattcgt tcaatccaaa
     4141 ctcgttgaaa tagttgaacg ttgcatcctc cttgaactct ttgaggtggt tcctcactgc
     4201 tgcaaatttt tcataggtgg atgccgtcca gttattctgg ttaccgcact cttttacaaa
     4261 ctcatcgaac acctcccaaa agctgacagg ggcttcttcc ggctgttctt cactggtgtc
     4321 tttcattctc atgttgaaag cttccttcaa ctgttgggtc gttggcatga cctcctgcac
     4381 ctcaaattcc ttgaaaatat tctggatttc ggcatagtat ttcagcaagt ccgtattgat
     4441 ttcggctgca ctttgcttta gcttgttggt acatccgttc tttacccgct gcttatctgc
     4501 atcccatttg gctacgtcaa tccggtagcc cgttgtaaac tcgatacgtt ggctggcaaa
     4561 gatgacacgc atacggatgg gtacgttctc tacgattggc acaccgttct ttttccggct
     4621 ctccaatgca aaaatgatgt tgcgcttgat attcataatt gggtgcgttt gaaattctac
     4681 acccaaatat acacccaatt attgagatag caaaagacat ttagaaacat ttacttttac
     4741 tctatattgt aatttacact tgattatcag tcgtttgcag ttttatgata ttctgtgaaa
     4801 gtataagttc gagagcctgt ctctccgcaa aaaacgctga aaatcagcag attgcaaaac
     4861 aaacaccctg ttttacaccc aagaatgtaa agtcgggtgt ttttgtttta tttaagataa
     4921 tacaaccact acataataaa agagtagcga tattaaaaga atccgatgag aaaagactaa
     4981 tatttatcta tccattcagt ttgatttctc aggactttac atcgtcctga aagtatttgt
     5041 tggtaccggt accgaggacg cgacagctgc agacgcgtta tgagtaaact tggtctgaca
     5101 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     5161 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5221 ccagtgctgc aatgataccg cgggacccac gctcaccggc tccagattta tcagcaataa
     5281 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5341 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5401 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5461 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5521 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5581 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5641 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5701 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5761 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5821 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5881 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5941 cacggaaatg ttgaatactc atactcttcc tttttcaatc atggccgcgg gattaaaagt
     6001 cggggattgg tgaacaaaaa ggtgtttctc tctttaagag aaatatcgtt ttgctaaaca
     6061 gttgatattg aggtatcatt ttatcgtaaa agacattttt gctcaacaat tgcttgacgg
     6121 aaatcaacaa attttagcat tttgtaaaaa agtcgctata taatttggtg aattggagtt
     6181 attttcatat ttttgcatcc cgaagagttt ctcttaaaga gagaaacatc ttttgcatac
     6241 cttttccgac cgaattttta tgtcgtaaag aggggctttg cagggggtgg actcagaaag
     6301 atgagaatag atgactattg tagttgaaac acatagaaag ttgctgatat acagaccgat
     6361 acgcatatcg ggatgaacca tgagtacgtt cttttctcaa aaaacataaa tattcgaaaa
     6421 gagatgcaat aaattaagga gaggttataa tgaacaaagt aaatataaaa gatagtcaaa
     6481 attttattac ttcaaaatat cacatagaaa aaataatgaa ttgcataagt ttagatgaaa
     6541 aagataacat ctttgaaata ggtgcaggga aaggtcattt tactgctgga ttggtaaaga
     6601 gatgtaattt tgtaacggcg atagaaattg attctaaatt atgtgaggta actcgtaata
     6661 agctcttaaa ttatcctaac tatcaaatag taaatgatga tatactgaaa tttacatttc
     6721 ctagccacaa tccatataaa atatttggca acatacctta caacataagc acaaatataa
     6781 ttcgaaaaat tgtttttgaa agttcagcca caataagtta tttaatagtg gaatatggtt
     6841 ttgctaaaat gttattagat acaaacagat cactagcatt gctgttaatg gcagaggtag
     6901 atatttctat attagcaaaa attcctaggt attatttcca tccaaaacct aaagtggata
     6961 gcacattaat tgtattaaaa agaaagccag caaaaatggc atttaaagag agaaaaaaat
     7021 atgaaacttt tgtaatgaaa tgggttaaca aagagtacga aaaactgttt acaaaaaatc
     7081 aatttaataa agctttaaaa catgcgagaa tatatgatat aaacaatatt agtttcgaac
     7141 aatttgtatc gctatttaat agttataaaa tatttaacgg ctaaaaacaa taggccacat
     7201 gcaactgtaa atgtttacgc gggtaccgac accgcggtgg aggggaattc ccatgtcagc
     7261 cgttaagtgt tcctgtgtca ctcaaaattg ctttgagagg ctctaagggc ttctcagtgc
     7321 gttacatccc tggcttgttg tccacaaccg ttaaacctta aaagctttaa aagccttata
     7381 tattcttttt tttcttataa aacttaaaac cttagaggct atttaagttg ctgatttata
     7441 ttaattttat tgttcaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtta
     7501 gccatgagag cttagtacgt tagccatgag ggtttagttc gttaaacatg agagcttagt
     7561 acgttaaaca tgagagctta gtacgtgaaa catgagagct tagtacgtac tatcaacagg
     7621 ttgaactgct gatcttcaga tcctctacgc cggacgcatc gtggccggat caattccgtt
     7681 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     7741 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     7801 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     7861 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     7921 ggctaccgcc ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag
     7981 gaagggcagc ccacctatca cggaattgat ccccctcgaa ttggaattct agcctggaat
     8041 agggaagcac cacatcacc
//
