LOCUS       Exported                8046 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 8046)
  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..8046
                     /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..1449
                     /label=PBT1004
     CDS             1450..2073
                     /codon_start=1
                     /label=TetR
                     /note="TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     terminator      2094..2122
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    2153..2260
                     /label=pTetR56
     protein_bind    2238..2256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2261..2342
                     /label=RBS1P-LP
     CDS             2343..3062
                     /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      3083..3110
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(3360..4643)
                     /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(5089..5949)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71346.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             6437..7171
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71348.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGNIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     misc_feature    7237..8010
                     /label=Origin
     rep_origin      7241..7629
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7800..7909
                     /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 aaccccgaat cagatgtgcg gagatatgct tggcatatct
     1021 ccgttttaat tgacataatc cgccggagta aagggatgaa tgatgcttac cgacgtgatt
     1081 taatcgtttt ttaataaatg aaatttattt tttcctgatt atacaaatag tacaaaagaa
     1141 tcttcttgtc tggtcttgtt ttttatattc cgcaacattt catcctttgt gtcatagtat
     1201 tagtcacctt cgaaacattt attgtttgta tccgacatgg atggagtttg gctagtcccc
     1261 aaatatgcgg atatttgtga agtgagattt cactatcaaa agtattggtc ctttcggacc
     1321 ggccgcctgt gaaggtagct ctattgtttg ttttgtttgt gtttggtgca tcttttttca
     1381 agcgagaaag aagatgcact atttatatct atctaccctg actatcggat attgaatgaa
     1441 aatttgccta tgtccagatt agataaaagt aaagtgatta acagcgcatt agagctgctt
     1501 aatgaggtcg gaatcgaagg tttaacaacc cgtaaactcg cccagaagct aggtgtagag
     1561 cagcctacat tgtattggca tgtaaaaaat aagcgggctt tgctcgacgc cttagccatt
     1621 gagatgttag ataggcacca tactcacttt tgccctttag aaggggaaag ctggcaagat
     1681 tttttacgta ataacgctaa aagttttaga tgtgctttac taagtcatcg cgatggagca
     1741 aaagtacatt taggtacacg gcctacagaa aaacagtatg aaactctcga aaatcaatta
     1801 gcctttttat gccaacaagg tttttcacta gagaatgcat tatatgcact cagcgctgtg
     1861 gggcatttta ctttaggttg cgtattggaa gatcaagagc atcaagtcgc taaagaagaa
     1921 agggaaacac ctactactga tagtatgccg ccattattac gacaagctat cgaattattt
     1981 gatcaccaag gtgcagagcc agccttctta ttcggccttg aattgatcat atgcggatta
     2041 gaaaaacaac ttaaatgtga aagtgggtct taatggccac atcggttcct gctagaaggc
     2101 ccatcctgac ggatggcctt ttggtattcg agcattacct catactttgg atttttgttt
     2161 tgtttgcaat ggttaatcta ttgttaaaat ttaaagtttc acttgaactt tcaaataatg
     2221 ttcttatatt tgcagtgtcc ctatcagtga tagagagtag gatcacacaa catttaaaaa
     2281 ataacattat gaaagcactc gaaaagagat tcaaaggtaa tattaacaat aatttatttt
     2341 caatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2401 acggcgacgt aaacggccac aagttcagcg tccgcggcga gggcgagggc gatgccacca
     2461 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2521 ccctcgtgac caccttaggc tacggcgtgg cctgcttcgc ccgctacccc gaccacatga
     2581 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2641 ctttcaagga cgacggtacc tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2701 tggtgaaccg catcgtgctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     2761 acaagctgga gtacaacttc aacagccaca aggtctatat cacggccgac aagcagaaga
     2821 acggcatcaa ggctaacttc aagacccgcc acaacgttga ggacggcggc gtgcagctcg
     2881 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2941 actacctgag ccatcagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg
     3001 tcctgaagga gagggtgacc gccgccggga ttacacatga catggacgag ctgtacaagt
     3061 aatagcgttc agctcgactg tgagaaggcc atcctgacgg atggcctttt ttttcgttat
     3121 aactagttgc gttcctagcc gctactagtg taattgccta tcttccagtg atggaacagc
     3181 atttgtgcat tggctgcaac aatcagcctt acttgtgcct gttctatttc cgaaccgacc
     3241 gcttgtatga atgcatcaaa attcgttttc tctacgttgg attccttgtt gctcatattg
     3301 tgatgataat ttctacaaat atagtcattg gtaactatct atgaaactgt ttgatacttt
     3361 tatagttgat taaacttgtt catggcattt gccttaatat catccgctat gtcaatgtag
     3421 ggtttcatag ctttgtagtc gctgtgtccc gtccatttca tgaccacctg tgccgggatt
     3481 ccgagagcca gcgcattgca gatgaatgtc ctttttcctg catgggtact gagcaaagcg
     3541 tatttgtgtg tgacttcatc aatacgttca tttcccttgt agtaggtttc ccgtacaggc
     3601 tcgttgattt ctgccagttc gcccagctct ttcaggtaat cgttcatctt ctggttgctg
     3661 atgacgggca gagccatgta attctcgaaa tggatgtcct tgtatttgtc cagtatggct
     3721 ttgctgtatt tgttcagttc aatcgtcagg ctgtcggcag tcttgactgt ggttatttcg
     3781 atgtggtcgg acttcacatc gcttcttttc agattgcgaa catccgaata ccgcaaactc
     3841 gtaaagcagc agaacaggaa aacatcacgc acacgttcca ggtattgctt atccttgggt
     3901 atctggtagt ctttcagctt gttcagttca tcccaagtca ggaagattac ttttttcgag
     3961 gtggttttca gtttcggttt gaacgtatcg tatgcaatgt tctgatgatg tcctttcttg
     4021 aagctccagc gcaggaacca tttgaggaat cccatttgct tgccgatggt gctgtttctc
     4081 atatccttgg tgtcacgcag gaagttgacg tattcgttca atccaaactc gttgaaatag
     4141 ttgaacgttg catcctcctt gaactctttg aggtggttcc tcactgctgc aaatttttca
     4201 taggtggatg ccgtccagtt attctggtta ccgcactctt ttacaaactc atcgaacacc
     4261 tcccaaaagc tgacaggggc ttcttccggc tgttcttcac tggtgtcttt cattctcatg
     4321 ttgaaagctt ccttcaactg ttgggtcgtt ggcatgacct cctgcacctc aaattccttg
     4381 aaaatattct ggatttcggc atagtatttc agcaagtccg tattgatttc ggctgcactt
     4441 tgctttagct tgttggtaca tccgttcttt acccgctgct tatctgcatc ccatttggct
     4501 acgtcaatcc ggtagcccgt tgtaaactcg atacgttggc tggcaaagat gacacgcata
     4561 cggatgggta cgttctctac gattggcaca ccgttctttt tccggctctc caatgcaaaa
     4621 atgatgttgc gcttgatatt cataattggg tgcgtttgaa attctacacc caaatataca
     4681 cccaattatt gagatagcaa aagacattta gaaacattta cttttactct atattgtaat
     4741 ttacacttga ttatcagtcg tttgcagttt tatgatattc tgtgaaagta taagttcgag
     4801 agcctgtctc tccgcaaaaa acgctgaaaa tcagcagatt gcaaaacaaa caccctgttt
     4861 tacacccaag aatgtaaagt cgggtgtttt tgttttattt aagataatac aaccactaca
     4921 taataaaaga gtagcgatat taaaagaatc cgatgagaaa agactaatat ttatctatcc
     4981 attcagtttg atttctcagg actttacatc gtcctgaaag tatttgttgg taccggtacc
     5041 gaggacgcga cagctgcaga cgcgttatga gtaaacttgg tctgacagtt accaatgctt
     5101 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5161 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5221 gataccgcgg gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5281 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5341 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5401 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5461 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5521 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5581 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5641 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5701 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5761 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5821 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5881 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5941 aatactcata ctcttccttt ttcaatcatg gccgcgggat taaaagtcgg ggattggtga
     6001 acaaaaaggt gtttctctct ttaagagaaa tatcgttttg ctaaacagtt gatattgagg
     6061 tatcatttta tcgtaaaaga catttttgct caacaattgc ttgacggaaa tcaacaaatt
     6121 ttagcatttt gtaaaaaagt cgctatataa tttggtgaat tggagttatt ttcatatttt
     6181 tgcatcccga agagtttctc ttaaagagag aaacatcttt tgcatacctt ttccgaccga
     6241 atttttatgt cgtaaagagg ggctttgcag ggggtggact cagaaagatg agaatagatg
     6301 actattgtag ttgaaacaca tagaaagttg ctgatataca gaccgatacg catatcggga
     6361 tgaaccatga gtacgttctt ttctcaaaaa acataaatat tcgaaaagag atgcaataaa
     6421 ttaaggagag gttataatga acaaagtaaa tataaaagat agtcaaaatt ttattacttc
     6481 aaaatatcac atagaaaaaa taatgaattg cataagttta gatgaaaaag ataacatctt
     6541 tgaaataggt gcagggaaag gtcattttac tgctggattg gtaaagagat gtaattttgt
     6601 aacggcgata gaaattgatt ctaaattatg tgaggtaact cgtaataagc tcttaaatta
     6661 tcctaactat caaatagtaa atgatgatat actgaaattt acatttccta gccacaatcc
     6721 atataaaata tttggcaaca taccttacaa cataagcaca aatataattc gaaaaattgt
     6781 ttttgaaagt tcagccacaa taagttattt aatagtggaa tatggttttg ctaaaatgtt
     6841 attagataca aacagatcac tagcattgct gttaatggca gaggtagata tttctatatt
     6901 agcaaaaatt cctaggtatt atttccatcc aaaacctaaa gtggatagca cattaattgt
     6961 attaaaaaga aagccagcaa aaatggcatt taaagagaga aaaaaatatg aaacttttgt
     7021 aatgaaatgg gttaacaaag agtacgaaaa actgtttaca aaaaatcaat ttaataaagc
     7081 tttaaaacat gcgagaatat atgatataaa caatattagt ttcgaacaat ttgtatcgct
     7141 atttaatagt tataaaatat ttaacggcta aaaacaatag gccacatgca actgtaaatg
     7201 tttacgcggg taccgacacc gcggtggagg ggaattccca tgtcagccgt taagtgttcc
     7261 tgtgtcactc aaaattgctt tgagaggctc taagggcttc tcagtgcgtt acatccctgg
     7321 cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag ccttatatat tctttttttt
     7381 cttataaaac ttaaaacctt agaggctatt taagttgctg atttatatta attttattgt
     7441 tcaaacatga gagcttagta cgtgaaacat gagagcttag tacgttagcc atgagagctt
     7501 agtacgttag ccatgagggt ttagttcgtt aaacatgaga gcttagtacg ttaaacatga
     7561 gagcttagta cgtgaaacat gagagcttag tacgtactat caacaggttg aactgctgat
     7621 cttcagatcc tctacgccgg acgcatcgtg gccggatcaa ttccgttttc cgctgcataa
     7681 ccctgcttcg gggtcattat agcgattttt tcggtatatc catccttttt cgcacgatat
     7741 acaggatttt gccaaagggt tcgtgtagac tttccttggt gtatccaacg gcgtcagccg
     7801 ggcaggatag gtgaagtagg cccacccgcg agcgggtgtt ccttcttcac tgtcccttat
     7861 tcgcacctgg cggtgctcaa cgggaatcct gctctgcgag gctggccggc taccgccggc
     7921 gtaacagatg agggcaagcg gatggctgat gaaaccaagc caaccaggaa gggcagccca
     7981 cctatcacgg aattgatccc cctcgaattg gaattctagc ctggaatagg gaagcaccac
     8041 atcacc
//
