LOCUS       Exported                7909 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 7909)
  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..7909
                     /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"
     misc_feature    900..954
                     /label=gibson backbone
     terminator      919..946
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        989..1312
                     /label=PBT0722
     CDS             1313..1936
                     /codon_start=1
                     /label=TetR
                     /note="TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     terminator      1957..1985
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    2016..2123
                     /label=pTetR56
     protein_bind    2101..2119
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2124..2205
                     /label=RBS1P-LP
     CDS             2206..2925
                     /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      2946..2973
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3223..4506)
                     /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(4952..5812)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71346.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             6300..7034
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71348.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGNIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     misc_feature    7100..7873
                     /label=Origin
     rep_origin      7104..7492
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7663..7772
                     /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 aaccccgact catggagtgg aaacattaac cgacaaatcc
     1021 tattaatcag attgtttgca caaataaaag tatgtgctct acacaacata gaaatttgac
     1081 tcaggcatcg aactggcaac taaataaaaa tgatataaac acataatata taaaggttaa
     1141 ggtctacata accaagtttt tactacattc tattcctagt ttcagctacc tttcccattc
     1201 taattattgc atatttattt tgaaaatatg caataagttc ttatctttgc acccgaatta
     1261 aagaaccaga agatgaatca aacattacaa ttacataacg cccttatatt tcatgtccag
     1321 attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga
     1381 aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg
     1441 gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca
     1501 ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc
     1561 taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac
     1621 acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca
     1681 aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg
     1741 ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac
     1801 tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga
     1861 gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg
     1921 tgaaagtggg tcttaatggc cacatcggtt cctgctagaa ggcccatcct gacggatggc
     1981 cttttggtat tcgagcatta cctcatactt tggatttttg ttttgtttgc aatggttaat
     2041 ctattgttaa aatttaaagt ttcacttgaa ctttcaaata atgttcttat atttgcagtg
     2101 tccctatcag tgatagagag taggatcaca caacatttaa aaaataacat tatgaaagca
     2161 ctcgaaaaga gattcaaagg taatattaac aataatttat tttcaatggt gagcaagggc
     2221 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2281 cacaagttca gcgtccgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
     2341 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccacctta
     2401 ggctacggcg tggcctgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2461 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tctctttcaa ggacgacggt
     2521 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgtg
     2581 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2641 ttcaacagcc acaaggtcta tatcacggcc gacaagcaga agaacggcat caaggctaac
     2701 ttcaagaccc gccacaacgt tgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     2761 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagccatcag
     2821 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgaa ggagagggtg
     2881 accgccgccg ggattacaca tgacatggac gagctgtaca agtaatagcg ttcagctcga
     2941 ctgtgagaag gccatcctga cggatggcct tttttttcgt tataactagt tgcgttccta
     3001 gccgctacta gtgtaattgc ctatcttcca gtgatggaac agcatttgtg cattggctgc
     3061 aacaatcagc cttacttgtg cctgttctat ttccgaaccg accgcttgta tgaatgcatc
     3121 aaaattcgtt ttctctacgt tggattcctt gttgctcata ttgtgatgat aatttctaca
     3181 aatatagtca ttggtaacta tctatgaaac tgtttgatac ttttatagtt gattaaactt
     3241 gttcatggca tttgccttaa tatcatccgc tatgtcaatg tagggtttca tagctttgta
     3301 gtcgctgtgt cccgtccatt tcatgaccac ctgtgccggg attccgagag ccagcgcatt
     3361 gcagatgaat gtcctttttc ctgcatgggt actgagcaaa gcgtatttgt gtgtgacttc
     3421 atcaatacgt tcatttccct tgtagtaggt ttcccgtaca ggctcgttga tttctgccag
     3481 ttcgcccagc tctttcaggt aatcgttcat cttctggttg ctgatgacgg gcagagccat
     3541 gtaattctcg aaatggatgt ccttgtattt gtccagtatg gctttgctgt atttgttcag
     3601 ttcaatcgtc aggctgtcgg cagtcttgac tgtggttatt tcgatgtggt cggacttcac
     3661 atcgcttctt ttcagattgc gaacatccga ataccgcaaa ctcgtaaagc agcagaacag
     3721 gaaaacatca cgcacacgtt ccaggtattg cttatccttg ggtatctggt agtctttcag
     3781 cttgttcagt tcatcccaag tcaggaagat tacttttttc gaggtggttt tcagtttcgg
     3841 tttgaacgta tcgtatgcaa tgttctgatg atgtcctttc ttgaagctcc agcgcaggaa
     3901 ccatttgagg aatcccattt gcttgccgat ggtgctgttt ctcatatcct tggtgtcacg
     3961 caggaagttg acgtattcgt tcaatccaaa ctcgttgaaa tagttgaacg ttgcatcctc
     4021 cttgaactct ttgaggtggt tcctcactgc tgcaaatttt tcataggtgg atgccgtcca
     4081 gttattctgg ttaccgcact cttttacaaa ctcatcgaac acctcccaaa agctgacagg
     4141 ggcttcttcc ggctgttctt cactggtgtc tttcattctc atgttgaaag cttccttcaa
     4201 ctgttgggtc gttggcatga cctcctgcac ctcaaattcc ttgaaaatat tctggatttc
     4261 ggcatagtat ttcagcaagt ccgtattgat ttcggctgca ctttgcttta gcttgttggt
     4321 acatccgttc tttacccgct gcttatctgc atcccatttg gctacgtcaa tccggtagcc
     4381 cgttgtaaac tcgatacgtt ggctggcaaa gatgacacgc atacggatgg gtacgttctc
     4441 tacgattggc acaccgttct ttttccggct ctccaatgca aaaatgatgt tgcgcttgat
     4501 attcataatt gggtgcgttt gaaattctac acccaaatat acacccaatt attgagatag
     4561 caaaagacat ttagaaacat ttacttttac tctatattgt aatttacact tgattatcag
     4621 tcgtttgcag ttttatgata ttctgtgaaa gtataagttc gagagcctgt ctctccgcaa
     4681 aaaacgctga aaatcagcag attgcaaaac aaacaccctg ttttacaccc aagaatgtaa
     4741 agtcgggtgt ttttgtttta tttaagataa tacaaccact acataataaa agagtagcga
     4801 tattaaaaga atccgatgag aaaagactaa tatttatcta tccattcagt ttgatttctc
     4861 aggactttac atcgtcctga aagtatttgt tggtaccggt accgaggacg cgacagctgc
     4921 agacgcgtta tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4981 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5041 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgggacccac
     5101 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5161 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5221 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5281 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5341 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5401 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5461 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5521 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5581 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5641 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5701 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5761 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5821 tttttcaatc atggccgcgg gattaaaagt cggggattgg tgaacaaaaa ggtgtttctc
     5881 tctttaagag aaatatcgtt ttgctaaaca gttgatattg aggtatcatt ttatcgtaaa
     5941 agacattttt gctcaacaat tgcttgacgg aaatcaacaa attttagcat tttgtaaaaa
     6001 agtcgctata taatttggtg aattggagtt attttcatat ttttgcatcc cgaagagttt
     6061 ctcttaaaga gagaaacatc ttttgcatac cttttccgac cgaattttta tgtcgtaaag
     6121 aggggctttg cagggggtgg actcagaaag atgagaatag atgactattg tagttgaaac
     6181 acatagaaag ttgctgatat acagaccgat acgcatatcg ggatgaacca tgagtacgtt
     6241 cttttctcaa aaaacataaa tattcgaaaa gagatgcaat aaattaagga gaggttataa
     6301 tgaacaaagt aaatataaaa gatagtcaaa attttattac ttcaaaatat cacatagaaa
     6361 aaataatgaa ttgcataagt ttagatgaaa aagataacat ctttgaaata ggtgcaggga
     6421 aaggtcattt tactgctgga ttggtaaaga gatgtaattt tgtaacggcg atagaaattg
     6481 attctaaatt atgtgaggta actcgtaata agctcttaaa ttatcctaac tatcaaatag
     6541 taaatgatga tatactgaaa tttacatttc ctagccacaa tccatataaa atatttggca
     6601 acatacctta caacataagc acaaatataa ttcgaaaaat tgtttttgaa agttcagcca
     6661 caataagtta tttaatagtg gaatatggtt ttgctaaaat gttattagat acaaacagat
     6721 cactagcatt gctgttaatg gcagaggtag atatttctat attagcaaaa attcctaggt
     6781 attatttcca tccaaaacct aaagtggata gcacattaat tgtattaaaa agaaagccag
     6841 caaaaatggc atttaaagag agaaaaaaat atgaaacttt tgtaatgaaa tgggttaaca
     6901 aagagtacga aaaactgttt acaaaaaatc aatttaataa agctttaaaa catgcgagaa
     6961 tatatgatat aaacaatatt agtttcgaac aatttgtatc gctatttaat agttataaaa
     7021 tatttaacgg ctaaaaacaa taggccacat gcaactgtaa atgtttacgc gggtaccgac
     7081 accgcggtgg aggggaattc ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg
     7141 ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg
     7201 ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac
     7261 cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta
     7321 gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag
     7381 ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa
     7441 catgagagct tagtacgtac tatcaacagg ttgaactgct gatcttcaga tcctctacgc
     7501 cggacgcatc gtggccggat caattccgtt ttccgctgca taaccctgct tcggggtcat
     7561 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     7621 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     7681 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     7741 caacgggaat cctgctctgc gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa
     7801 gcggatggct gatgaaacca agccaaccag gaagggcagc ccacctatca cggaattgat
     7861 ccccctcgaa ttggaattct agcctggaat agggaagcac cacatcacc
//
