LOCUS       Exported                7614 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 7614)
  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..7614
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..120
                     /label=Promoter P_BfP1E6
     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"
     misc_feature    989..1017
                     /label=promoter insertion site
     CDS             1018..1641
                     /codon_start=1
                     /label=TetR
                     /note="TetR"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     terminator      1662..1690
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_feature    1721..1828
                     /label=pTetR56
     protein_bind    1806..1824
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1829..1910
                     /label=RBS1P-LP
     CDS             1911..2630
                     /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      2651..2678
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2928..4211)
                     /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(4657..5517)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71346.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             6005..6739
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71348.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGNIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     rep_origin      6809..7197
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7368..7477
                     /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 aaccccgaag agaccgcgga tgtacgaagt ggtctcgatg
     1021 tccagattag ataaaagtaa agtgattaac agcgcattag agctgcttaa tgaggtcgga
     1081 atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca gcctacattg
     1141 tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga gatgttagat
     1201 aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt tttacgtaat
     1261 aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa agtacattta
     1321 ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc ctttttatgc
     1381 caacaaggtt tttcactaga gaatgcatta tatgcactca gcgctgtggg gcattttact
     1441 ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag ggaaacacct
     1501 actactgata gtatgccgcc attattacga caagctatcg aattatttga tcaccaaggt
     1561 gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga aaaacaactt
     1621 aaatgtgaaa gtgggtctta atggccacat cggttcctgc tagaaggccc atcctgacgg
     1681 atggcctttt ggtattcgag cattacctca tactttggat ttttgttttg tttgcaatgg
     1741 ttaatctatt gttaaaattt aaagtttcac ttgaactttc aaataatgtt cttatatttg
     1801 cagtgtccct atcagtgata gagagtagga tcacacaaca tttaaaaaat aacattatga
     1861 aagcactcga aaagagattc aaaggtaata ttaacaataa tttattttca atggtgagca
     1921 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     1981 acggccacaa gttcagcgtc cgcggcgagg gcgagggcga tgccaccaac ggcaagctga
     2041 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2101 ccttaggcta cggcgtggcc tgcttcgccc gctaccccga ccacatgaag cagcacgact
     2161 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctct ttcaaggacg
     2221 acggtaccta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2281 tcgtgctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2341 acaacttcaa cagccacaag gtctatatca cggccgacaa gcagaagaac ggcatcaagg
     2401 ctaacttcaa gacccgccac aacgttgagg acggcggcgt gcagctcgcc gaccactacc
     2461 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcc
     2521 atcagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgaaggaga
     2581 gggtgaccgc cgccgggatt acacatgaca tggacgagct gtacaagtaa tagcgttcag
     2641 ctcgactgtg agaaggccat cctgacggat ggcctttttt ttcgttataa ctagttgcgt
     2701 tcctagccgc tactagtgta attgcctatc ttccagtgat ggaacagcat ttgtgcattg
     2761 gctgcaacaa tcagccttac ttgtgcctgt tctatttccg aaccgaccgc ttgtatgaat
     2821 gcatcaaaat tcgttttctc tacgttggat tccttgttgc tcatattgtg atgataattt
     2881 ctacaaatat agtcattggt aactatctat gaaactgttt gatactttta tagttgatta
     2941 aacttgttca tggcatttgc cttaatatca tccgctatgt caatgtaggg tttcatagct
     3001 ttgtagtcgc tgtgtcccgt ccatttcatg accacctgtg ccgggattcc gagagccagc
     3061 gcattgcaga tgaatgtcct ttttcctgca tgggtactga gcaaagcgta tttgtgtgtg
     3121 acttcatcaa tacgttcatt tcccttgtag taggtttccc gtacaggctc gttgatttct
     3181 gccagttcgc ccagctcttt caggtaatcg ttcatcttct ggttgctgat gacgggcaga
     3241 gccatgtaat tctcgaaatg gatgtccttg tatttgtcca gtatggcttt gctgtatttg
     3301 ttcagttcaa tcgtcaggct gtcggcagtc ttgactgtgg ttatttcgat gtggtcggac
     3361 ttcacatcgc ttcttttcag attgcgaaca tccgaatacc gcaaactcgt aaagcagcag
     3421 aacaggaaaa catcacgcac acgttccagg tattgcttat ccttgggtat ctggtagtct
     3481 ttcagcttgt tcagttcatc ccaagtcagg aagattactt ttttcgaggt ggttttcagt
     3541 ttcggtttga acgtatcgta tgcaatgttc tgatgatgtc ctttcttgaa gctccagcgc
     3601 aggaaccatt tgaggaatcc catttgcttg ccgatggtgc tgtttctcat atccttggtg
     3661 tcacgcagga agttgacgta ttcgttcaat ccaaactcgt tgaaatagtt gaacgttgca
     3721 tcctccttga actctttgag gtggttcctc actgctgcaa atttttcata ggtggatgcc
     3781 gtccagttat tctggttacc gcactctttt acaaactcat cgaacacctc ccaaaagctg
     3841 acaggggctt cttccggctg ttcttcactg gtgtctttca ttctcatgtt gaaagcttcc
     3901 ttcaactgtt gggtcgttgg catgacctcc tgcacctcaa attccttgaa aatattctgg
     3961 atttcggcat agtatttcag caagtccgta ttgatttcgg ctgcactttg ctttagcttg
     4021 ttggtacatc cgttctttac ccgctgctta tctgcatccc atttggctac gtcaatccgg
     4081 tagcccgttg taaactcgat acgttggctg gcaaagatga cacgcatacg gatgggtacg
     4141 ttctctacga ttggcacacc gttctttttc cggctctcca atgcaaaaat gatgttgcgc
     4201 ttgatattca taattgggtg cgtttgaaat tctacaccca aatatacacc caattattga
     4261 gatagcaaaa gacatttaga aacatttact tttactctat attgtaattt acacttgatt
     4321 atcagtcgtt tgcagtttta tgatattctg tgaaagtata agttcgagag cctgtctctc
     4381 cgcaaaaaac gctgaaaatc agcagattgc aaaacaaaca ccctgtttta cacccaagaa
     4441 tgtaaagtcg ggtgtttttg ttttatttaa gataatacaa ccactacata ataaaagagt
     4501 agcgatatta aaagaatccg atgagaaaag actaatattt atctatccat tcagtttgat
     4561 ttctcaggac tttacatcgt cctgaaagta tttgttggta ccggtaccga ggacgcgaca
     4621 gctgcagacg cgttatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4681 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4741 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcggga
     4801 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4861 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4921 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4981 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5041 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5101 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5161 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5221 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5281 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5341 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5401 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5461 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5521 cttccttttt caatcatggc cgcgggatta aaagtcgggg attggtgaac aaaaaggtgt
     5581 ttctctcttt aagagaaata tcgttttgct aaacagttga tattgaggta tcattttatc
     5641 gtaaaagaca tttttgctca acaattgctt gacggaaatc aacaaatttt agcattttgt
     5701 aaaaaagtcg ctatataatt tggtgaattg gagttatttt catatttttg catcccgaag
     5761 agtttctctt aaagagagaa acatcttttg catacctttt ccgaccgaat ttttatgtcg
     5821 taaagagggg ctttgcaggg ggtggactca gaaagatgag aatagatgac tattgtagtt
     5881 gaaacacata gaaagttgct gatatacaga ccgatacgca tatcgggatg aaccatgagt
     5941 acgttctttt ctcaaaaaac ataaatattc gaaaagagat gcaataaatt aaggagaggt
     6001 tataatgaac aaagtaaata taaaagatag tcaaaatttt attacttcaa aatatcacat
     6061 agaaaaaata atgaattgca taagtttaga tgaaaaagat aacatctttg aaataggtgc
     6121 agggaaaggt cattttactg ctggattggt aaagagatgt aattttgtaa cggcgataga
     6181 aattgattct aaattatgtg aggtaactcg taataagctc ttaaattatc ctaactatca
     6241 aatagtaaat gatgatatac tgaaatttac atttcctagc cacaatccat ataaaatatt
     6301 tggcaacata ccttacaaca taagcacaaa tataattcga aaaattgttt ttgaaagttc
     6361 agccacaata agttatttaa tagtggaata tggttttgct aaaatgttat tagatacaaa
     6421 cagatcacta gcattgctgt taatggcaga ggtagatatt tctatattag caaaaattcc
     6481 taggtattat ttccatccaa aacctaaagt ggatagcaca ttaattgtat taaaaagaaa
     6541 gccagcaaaa atggcattta aagagagaaa aaaatatgaa acttttgtaa tgaaatgggt
     6601 taacaaagag tacgaaaaac tgtttacaaa aaatcaattt aataaagctt taaaacatgc
     6661 gagaatatat gatataaaca atattagttt cgaacaattt gtatcgctat ttaatagtta
     6721 taaaatattt aacggctaaa aacaataggc cacatgcaac tgtaaatgtt tacgcgggta
     6781 ccgacaccgc ggtggagggg aattcccatg tcagccgtta agtgttcctg tgtcactcaa
     6841 aattgctttg agaggctcta agggcttctc agtgcgttac atccctggct tgttgtccac
     6901 aaccgttaaa ccttaaaagc tttaaaagcc ttatatattc ttttttttct tataaaactt
     6961 aaaaccttag aggctattta agttgctgat ttatattaat tttattgttc aaacatgaga
     7021 gcttagtacg tgaaacatga gagcttagta cgttagccat gagagcttag tacgttagcc
     7081 atgagggttt agttcgttaa acatgagagc ttagtacgtt aaacatgaga gcttagtacg
     7141 tgaaacatga gagcttagta cgtactatca acaggttgaa ctgctgatct tcagatcctc
     7201 tacgccggac gcatcgtggc cggatcaatt ccgttttccg ctgcataacc ctgcttcggg
     7261 gtcattatag cgattttttc ggtatatcca tcctttttcg cacgatatac aggattttgc
     7321 caaagggttc gtgtagactt tccttggtgt atccaacggc gtcagccggg caggataggt
     7381 gaagtaggcc cacccgcgag cgggtgttcc ttcttcactg tcccttattc gcacctggcg
     7441 gtgctcaacg ggaatcctgc tctgcgaggc tggccggcta ccgccggcgt aacagatgag
     7501 ggcaagcgga tggctgatga aaccaagcca accaggaagg gcagcccacc tatcacggaa
     7561 ttgatccccc tcgaattgga attctagcct ggaataggga agcaccacat cacc
//
