LOCUS       Exported                8083 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 8083)
  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..8083
                     /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..1486
                     /label=PBT0981
     CDS             1487..2110
                     /codon_start=1
                     /label=TetR
                     /note="TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     terminator      2131..2159
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    2190..2297
                     /label=pTetR56
     protein_bind    2275..2293
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2298..2379
                     /label=RBS1P-LP
     CDS             2380..3099
                     /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      3120..3147
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3397..4680)
                     /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(5126..5986)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71346.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             6474..7208
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71348.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGNIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     misc_feature    7274..8047
                     /label=Origin
     rep_origin      7278..7666
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7837..7946
                     /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 aaccccgaaa gttctcacgg aaagggaata atggtgatat
     1021 atatatacat aaaccttaaa caggctctgt ttttatatcc tatcatgaaa tatttcccga
     1081 acatagtggg aaccggtagc aagaatcgtg tttctaaaaa agcaaatagg cgttacaaag
     1141 cttatagatg tgtttttata ggtttattta ccccttttgt tctttttgaa aagcctaata
     1201 ctggattttt agactttact ggaatttgag aatgaaaaat gaggtagttt agatcgaaat
     1261 acagcattta atgtaagtac ttaaatatga cattcgcttt tgttgaatct ttgcttcgga
     1321 tcattgcttt ggggctgttt ccagttcagc tttagcactg aaattctcgc gaaatttcaa
     1381 tgaatatctt cgatgataaa aaagataagt tataaaattc agtttctttt ttaaagataa
     1441 aatagtatct ttgatgcaag attcatttcc actaatataa tatacaatgt ccagattaga
     1501 taaaagtaaa gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt
     1561 aacaacccgt aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt
     1621 aaaaaataag cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac
     1681 tcacttttgc cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag
     1741 ttttagatgt gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc
     1801 tacagaaaaa cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt
     1861 ttcactagag aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt
     1921 attggaagat caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag
     1981 tatgccgcca ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc
     2041 cttcttattc ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag
     2101 tgggtcttaa tggccacatc ggttcctgct agaaggccca tcctgacgga tggccttttg
     2161 gtattcgagc attacctcat actttggatt tttgttttgt ttgcaatggt taatctattg
     2221 ttaaaattta aagtttcact tgaactttca aataatgttc ttatatttgc agtgtcccta
     2281 tcagtgatag agagtaggat cacacaacat ttaaaaaata acattatgaa agcactcgaa
     2341 aagagattca aaggtaatat taacaataat ttattttcaa tggtgagcaa gggcgaggag
     2401 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     2461 ttcagcgtcc gcggcgaggg cgagggcgat gccaccaacg gcaagctgac cctgaagttc
     2521 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttaggctac
     2581 ggcgtggcct gcttcgcccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     2641 gccatgcccg aaggctacgt ccaggagcgc accatctctt tcaaggacga cggtacctac
     2701 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgtgctgaag
     2761 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caacttcaac
     2821 agccacaagg tctatatcac ggccgacaag cagaagaacg gcatcaaggc taacttcaag
     2881 acccgccaca acgttgagga cggcggcgtg cagctcgccg accactacca gcagaacacc
     2941 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcca tcagtccaag
     3001 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgaaggagag ggtgaccgcc
     3061 gccgggatta cacatgacat ggacgagctg tacaagtaat agcgttcagc tcgactgtga
     3121 gaaggccatc ctgacggatg gccttttttt tcgttataac tagttgcgtt cctagccgct
     3181 actagtgtaa ttgcctatct tccagtgatg gaacagcatt tgtgcattgg ctgcaacaat
     3241 cagccttact tgtgcctgtt ctatttccga accgaccgct tgtatgaatg catcaaaatt
     3301 cgttttctct acgttggatt ccttgttgct catattgtga tgataatttc tacaaatata
     3361 gtcattggta actatctatg aaactgtttg atacttttat agttgattaa acttgttcat
     3421 ggcatttgcc ttaatatcat ccgctatgtc aatgtagggt ttcatagctt tgtagtcgct
     3481 gtgtcccgtc catttcatga ccacctgtgc cgggattccg agagccagcg cattgcagat
     3541 gaatgtcctt tttcctgcat gggtactgag caaagcgtat ttgtgtgtga cttcatcaat
     3601 acgttcattt cccttgtagt aggtttcccg tacaggctcg ttgatttctg ccagttcgcc
     3661 cagctctttc aggtaatcgt tcatcttctg gttgctgatg acgggcagag ccatgtaatt
     3721 ctcgaaatgg atgtccttgt atttgtccag tatggctttg ctgtatttgt tcagttcaat
     3781 cgtcaggctg tcggcagtct tgactgtggt tatttcgatg tggtcggact tcacatcgct
     3841 tcttttcaga ttgcgaacat ccgaataccg caaactcgta aagcagcaga acaggaaaac
     3901 atcacgcaca cgttccaggt attgcttatc cttgggtatc tggtagtctt tcagcttgtt
     3961 cagttcatcc caagtcagga agattacttt tttcgaggtg gttttcagtt tcggtttgaa
     4021 cgtatcgtat gcaatgttct gatgatgtcc tttcttgaag ctccagcgca ggaaccattt
     4081 gaggaatccc atttgcttgc cgatggtgct gtttctcata tccttggtgt cacgcaggaa
     4141 gttgacgtat tcgttcaatc caaactcgtt gaaatagttg aacgttgcat cctccttgaa
     4201 ctctttgagg tggttcctca ctgctgcaaa tttttcatag gtggatgccg tccagttatt
     4261 ctggttaccg cactctttta caaactcatc gaacacctcc caaaagctga caggggcttc
     4321 ttccggctgt tcttcactgg tgtctttcat tctcatgttg aaagcttcct tcaactgttg
     4381 ggtcgttggc atgacctcct gcacctcaaa ttccttgaaa atattctgga tttcggcata
     4441 gtatttcagc aagtccgtat tgatttcggc tgcactttgc tttagcttgt tggtacatcc
     4501 gttctttacc cgctgcttat ctgcatccca tttggctacg tcaatccggt agcccgttgt
     4561 aaactcgata cgttggctgg caaagatgac acgcatacgg atgggtacgt tctctacgat
     4621 tggcacaccg ttctttttcc ggctctccaa tgcaaaaatg atgttgcgct tgatattcat
     4681 aattgggtgc gtttgaaatt ctacacccaa atatacaccc aattattgag atagcaaaag
     4741 acatttagaa acatttactt ttactctata ttgtaattta cacttgatta tcagtcgttt
     4801 gcagttttat gatattctgt gaaagtataa gttcgagagc ctgtctctcc gcaaaaaacg
     4861 ctgaaaatca gcagattgca aaacaaacac cctgttttac acccaagaat gtaaagtcgg
     4921 gtgtttttgt tttatttaag ataatacaac cactacataa taaaagagta gcgatattaa
     4981 aagaatccga tgagaaaaga ctaatattta tctatccatt cagtttgatt tctcaggact
     5041 ttacatcgtc ctgaaagtat ttgttggtac cggtaccgag gacgcgacag ctgcagacgc
     5101 gttatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5161 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5221 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgggac ccacgctcac
     5281 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5341 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5401 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5461 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5521 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5581 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5641 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5701 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     5761 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5821 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5881 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5941 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6001 aatcatggcc gcgggattaa aagtcgggga ttggtgaaca aaaaggtgtt tctctcttta
     6061 agagaaatat cgttttgcta aacagttgat attgaggtat cattttatcg taaaagacat
     6121 ttttgctcaa caattgcttg acggaaatca acaaatttta gcattttgta aaaaagtcgc
     6181 tatataattt ggtgaattgg agttattttc atatttttgc atcccgaaga gtttctctta
     6241 aagagagaaa catcttttgc ataccttttc cgaccgaatt tttatgtcgt aaagaggggc
     6301 tttgcagggg gtggactcag aaagatgaga atagatgact attgtagttg aaacacatag
     6361 aaagttgctg atatacagac cgatacgcat atcgggatga accatgagta cgttcttttc
     6421 tcaaaaaaca taaatattcg aaaagagatg caataaatta aggagaggtt ataatgaaca
     6481 aagtaaatat aaaagatagt caaaatttta ttacttcaaa atatcacata gaaaaaataa
     6541 tgaattgcat aagtttagat gaaaaagata acatctttga aataggtgca gggaaaggtc
     6601 attttactgc tggattggta aagagatgta attttgtaac ggcgatagaa attgattcta
     6661 aattatgtga ggtaactcgt aataagctct taaattatcc taactatcaa atagtaaatg
     6721 atgatatact gaaatttaca tttcctagcc acaatccata taaaatattt ggcaacatac
     6781 cttacaacat aagcacaaat ataattcgaa aaattgtttt tgaaagttca gccacaataa
     6841 gttatttaat agtggaatat ggttttgcta aaatgttatt agatacaaac agatcactag
     6901 cattgctgtt aatggcagag gtagatattt ctatattagc aaaaattcct aggtattatt
     6961 tccatccaaa acctaaagtg gatagcacat taattgtatt aaaaagaaag ccagcaaaaa
     7021 tggcatttaa agagagaaaa aaatatgaaa cttttgtaat gaaatgggtt aacaaagagt
     7081 acgaaaaact gtttacaaaa aatcaattta ataaagcttt aaaacatgcg agaatatatg
     7141 atataaacaa tattagtttc gaacaatttg tatcgctatt taatagttat aaaatattta
     7201 acggctaaaa acaataggcc acatgcaact gtaaatgttt acgcgggtac cgacaccgcg
     7261 gtggagggga attcccatgt cagccgttaa gtgttcctgt gtcactcaaa attgctttga
     7321 gaggctctaa gggcttctca gtgcgttaca tccctggctt gttgtccaca accgttaaac
     7381 cttaaaagct ttaaaagcct tatatattct tttttttctt ataaaactta aaaccttaga
     7441 ggctatttaa gttgctgatt tatattaatt ttattgttca aacatgagag cttagtacgt
     7501 gaaacatgag agcttagtac gttagccatg agagcttagt acgttagcca tgagggttta
     7561 gttcgttaaa catgagagct tagtacgtta aacatgagag cttagtacgt gaaacatgag
     7621 agcttagtac gtactatcaa caggttgaac tgctgatctt cagatcctct acgccggacg
     7681 catcgtggcc ggatcaattc cgttttccgc tgcataaccc tgcttcgggg tcattatagc
     7741 gattttttcg gtatatccat cctttttcgc acgatataca ggattttgcc aaagggttcg
     7801 tgtagacttt ccttggtgta tccaacggcg tcagccgggc aggataggtg aagtaggccc
     7861 acccgcgagc gggtgttcct tcttcactgt cccttattcg cacctggcgg tgctcaacgg
     7921 gaatcctgct ctgcgaggct ggccggctac cgccggcgta acagatgagg gcaagcggat
     7981 ggctgatgaa accaagccaa ccaggaaggg cagcccacct atcacggaat tgatccccct
     8041 cgaattggaa ttctagcctg gaatagggaa gcaccacatc acc
//
