LOCUS       Exported                6708 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 6708)
  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..6708
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        12..54
                     /label=J23100
                     /note="J23100"
                     /note="J23100 standard promoter"
     RBS             55..85
                     /label=PET-RBS
                     /note="PET-RBS"
                     /note="pET3a RBS"
     CDS             86..796
                     /codon_start=1
                     /transl_table=11
                     /product="mCherry"
                     /label=mCherry
                     /protein_id="ARI71363.1"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     terminator      816..843
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             964..1677
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1684..1755
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1759..1776
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1799..1826
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2058..3341)
                     /codon_start=1
                     /transl_table=11
                     /product="NBU integrase"
                     /label=NBU integrase
                     /protein_id="ARI71364.1"
                     /translation="MNIKRNIIFALESRKKNGVPIVENVPIRMRVIFASQRIEFTTGYR
                     IDVAKWDADKQRVKSGCTNKLKQSAAEINTDLLKYYAEIQNIFKEFEVQEVMPTTQQLK
                     EAFNMRMKDTSEEQPEEAPVSFWEVFDEFVKECGNQNNWTASTYEKFAAVRNHLKEFKE
                     DATFNYFNEFGLNEYVNFLRDTKDMRNSTIGKQMGFLKWFLRWSFKKGHHQNIAYDTFK
                     PKLKTTSKKVIFLTWDELNKLKDYQIPKDKQYLERVRDVFLFCCFTSLRYSDVRNLKRS
                     DVKSDHIEITTVKTADSLTIELNKYSKAILDKYKDIHFENYMALPVISNQKMNDYLKEL
                     GELAEINEPVRETYYKGNERIDEVTPKYALLSTHAGRRTFICNALALGIPAQVVMKWTG
                     HSDYKAMKPYIDIADDIKANAMNKFNQL"
     CDS             complement(3781..4641)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71365.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             5129..5863
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71367.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     rep_origin      5929..6702
     rep_origin      5933..6321
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6492..6601
                     /note="incP origin of transfer"
ORIGIN
        1 gttctgagac cctatttgac ggctagctca gtcctaggta cagtgctagc gtacgtactt
       61 taactttaag aaggagatat acataatggt ttcgaaaggg gaagaagata acatggctat
      121 cattaaggag tttatgcgct tcaaggtgca catggagggc tctgtcaacg gccacgagtt
      181 cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt actcagaccg caaagttgaa
      241 agttacaaaa ggaggtccgt tgcctttcgc ttgggacatt ctgagcccgc aatttatgta
      301 cgggagcaaa gcctatgtta aacatccggc ggatattccc gattacctga aactgtcatt
      361 cccggagggg ttcaagtggg aacgcgtcat gaactttgaa gacggtggcg tggttacagt
      421 gactcaggat tcgtcccttc aagatggcga atttatctac aaagtcaaac tgcgcgggac
      481 aaattttccg agtgacggtc ctgttatgca gaagaagacc atggggtggg aagcaagctc
      541 cgaacgcatg taccccgagg acggtgcact caaaggagaa atcaagcagc gtctgaaact
      601 gaaagatggc gggcactacg atgcagaagt gaaaaccacc tataaggcga aaaagcctgt
      661 acaactgccg ggtgcctata atgtgaacat taaactggat ataacgagtc acaacgaaga
      721 ctatacaatc gtcgagcagt atgagcgtgc ggaaggacgt cattccactg gtggaatgga
      781 tgaactgtat aagtaaatcc tagctgatta gctcgagaag gccatcctga cggatggcct
      841 tttttttgta tcgagacgac tagtgtaatt gcctatcttc cagtgatgga acagcatttg
      901 tgcattggct gcaacaatca gccttacttg tgcctgttct atttccgaac cgaccgggtc
      961 tcaatgcgta aaggcgaaga gctgttcact ggtgtcgtcc ctattctggt ggaactggat
     1021 ggtgatgtca acggtcataa gttttccgtg cgtggcgagg gtgaaggtga cgcaactaat
     1081 ggtaaactga cgctgaagtt catctgtact actggtaaac tgccggttcc ttggccgact
     1141 ctggtaacga cgctgactta tggtgttcag tgctttgctc gttatccgga ccatatgaag
     1201 cagcatgact tcttcaagtc cgccatgccg gaaggctatg tgcaggaacg cacgatttcc
     1261 tttaaggatg acggcacgta caaaacgcgt gcggaagtga aatttgaagg cgataccctg
     1321 gtaaaccgca ttgagctgaa aggcattgac tttaaagaag acggcaatat cctgggccat
     1381 aagctggaat acaattttaa cagccacaat gtttacatca ccgccgataa acaaaaaaat
     1441 ggcattaaag cgaattttaa aattcgccac aacgtggagg atggcagcgt gcagctggct
     1501 gatcactacc agcaaaacac tccaatcggt gatggtcctg ttctgctgcc agacaatcac
     1561 tatctgagca cgcaaagcgt tctgtctaaa gatccgaacg agaaacgcga tcatatggtt
     1621 ctgctggagt tcgtaaccgc agcgggcatc acgcatggta tggatgaact gtacaaagga
     1681 tctgattaca aggacgatga tgataaggac tacaaagatg atgatgacaa agattataag
     1741 gacgatgatg ataaaggtca tcatcatcat catcactaaa tcctagctga ttagctcgag
     1801 aaggccatcc tgacggatgg cctttttttt gtatcgagac gactagtgta attgcctatc
     1861 ttccagtgat ggaacagcat ttgtgcattg gctgcaacaa tcagccttac ttgtgcctgt
     1921 tctatttccg aaccgaccgc ttgtatgaat ccatcaaaat tcgttttctc tatgttggat
     1981 tccttgttgc tcatattgtg atgataattt ctacaaatat agtcattggt aactatctat
     2041 gaaactgttt gatactttta tagttgatta aacttgttca tggcatttgc cttaatatca
     2101 tccgctatgt caatgtaggg tttcatagct ttgtagtcgc tgtgtcccgt ccatttcatg
     2161 accacctgtg ccgggattcc gagagccagc gcattgcaga tgaatgtcct tcttcctgca
     2221 tgggtactga gcaaagcgta tttgggtgtg acttcatcaa tacgttcatt tcccttgtag
     2281 taggtttccc gtacaggctc gttgatttct gccagttcgc ccagctcttt caggtaatcg
     2341 ttcatcttct ggttgctgat gacgggcaga gccatgtaat tctcgaaatg gatgtccttg
     2401 tatttgtcca gtatggcttt gctgtatttg ttcagttcaa tcgtcaggct gtcggcagtc
     2461 ttgactgtgg ttatttcgat gtggtcggac ttcacatcgc ttcttttcag attgcgaaca
     2521 tccgaatacc gcaaactcgt aaagcagcag aacaggaaaa catcacgcac acgttccagg
     2581 tattgcttat ccttgggtat ctggtagtct ttcagcttgt tcagttcatc ccaagtcagg
     2641 aagattactt ttttcgaggt ggttttcagt ttcggtttga acgtatcgta tgcaatgttc
     2701 tgatgatgtc ctttcttgaa gctccagcgc aggaaccatt tgaggaatcc catttgcttg
     2761 ccgatggtgc tgtttctcat atccttggtg tcacgcagga agttgacgta ttcgttcaat
     2821 ccaaactcgt tgaaatagtt gaacgttgca tcctccttga actctttgag gtggttcctc
     2881 actgctgcaa atttttcata ggtggatgcc gtccagttat tctggttacc gcactctttt
     2941 acaaactcat cgaacacctc ccaaaagctg acaggggctt cttccggctg ttcttcgctg
     3001 gtgtctttca ttctcatgtt gaaagcttcc ttcaactgtt gggtcgttgg catgacctcc
     3061 tgcacctcaa attccttgaa aatattctgg atttcggcat agtatttcag caagtccgta
     3121 ttgatttcgg ctgcactttg ctttagcttg ttggtacatc cgctctttac ccgctgctta
     3181 tctgcatccc atttggctac gtcaatccgg tagcccgttg taaactcgat gcgttggctg
     3241 gcaaagatga cacgcatacg gatgggtacg ttctctacga ttggcacacc gttctttttc
     3301 cggctctcca atgcaaaaat gatgttgcgc ttgatattca taattgggtg cgtttgaaat
     3361 tctacaccca aatatacacc caattattga gatagcaaaa gacatttaga aacatttact
     3421 tttactctat attgtaattt acacttgatt atcagtcgtt tgcagtctta tgatattctg
     3481 tgaaagtata agttcgagag cctgtctctc cgcaaaaaac gctgaaaatc agcagattgc
     3541 aaaacaaaca ccctgtttta cacccaagaa tgtaaagtcg gctgtttttg ttttatttaa
     3601 gataatacaa ccactacata ataaaagagt agcgatatta aaagaatccg atgagaaaag
     3661 actaatattt atctatccat tcagtttgat ttttcaggac tttacatcgt cctgaaagta
     3721 tttgttggta ccgaggacgc gacagctgca gacgcgttat gagtaaactt ggtctgacag
     3781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3901 cagtgctgca atgataccgc gggacccacg ctcaccggct ccagatttat cagcaataaa
     3961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     4201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     4261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4621 acggaaatgt tgaatactca tactcttcct ttttcaatca tggccgcggg attaaaagtc
     4681 ggggattggt gaacaaaaag gtgtttctct ctttaagaga aatatcgttt tgctaaacag
     4741 ttgatattga ggtatcattt tatcgtaaaa gacatttttg ctcaacaatt gcttgacgga
     4801 aatcaacaaa ttttagcatt ttgtaaaaaa gtcgctatat aatttggtga attggagtta
     4861 ttttcatatt tttgcatccc gaagagtttc tcttaaagag agaaacatct tttgcatacc
     4921 ttttccgacc gaatttttat gtcgtaaaga ggggctttgc agggggtgga ctcagaaaga
     4981 tgagaataga tgactattgt agttgaaaca catagaaagt tgctgatata cagaccgata
     5041 cgcatatcgg gatgaaccat gagtacgttc ttttctcaaa aaacataaat attcgaaaag
     5101 agatgcaata aattaaggag aggttataat gaacaaagta aatataaaag atagtcaaaa
     5161 ttttattact tcaaaatatc acatagaaaa aataatgaat tgcataagtt tagatgaaaa
     5221 agataacatc tttgaaatag gtgcagggaa aggtcatttt actgctggat tggtaaagag
     5281 atgtaatttt gtaacggcga tagaaattga ttctaaatta tgtgaggtaa ctcgtaataa
     5341 gctcttaaat tatcctaact atcaaatagt aaatgatgat atactgaaat ttacatttcc
     5401 tagccacaat ccatataaaa tatttggcag cataccttac aacataagca caaatataat
     5461 tcgaaaaatt gtttttgaaa gttcagccac aataagttat ttaatagtgg aatatggttt
     5521 tgctaaaatg ttattagata caaacagatc actagcattg ctgttaatgg cagaggtaga
     5581 tatttctata ttagcaaaaa ttcctaggta ttatttccat ccaaaaccta aagtggatag
     5641 cacattaatt gtattaaaaa gaaagccagc aaaaatggca tttaaagaga gaaaaaaata
     5701 tgaaactttt gtaatgaaat gggttaacaa agagtacgaa aaactgttta caaaaaatca
     5761 atttaataaa gctttaaaac atgcgagaat atatgatata aacaatatta gtttcgaaca
     5821 atttgtatcg ctatttaata gttataaaat atttaacggc taaaaacaat aggccacatg
     5881 caactgtaaa tgtttacgcg ggtaccgaca ccgcggtgga ggggaattcc catgtcagcc
     5941 gttaagtgtt cctgtgtcac tcaaaattgc tttgagaggc tctaagggct tctcagtgcg
     6001 ttacatccct ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat
     6061 attctttttt ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat
     6121 taattttatt gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag
     6181 ccatgagagc ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta
     6241 cgttaaacat gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt
     6301 tgaactgctg atcttcagat cctctacgcc ggacgcatcg tggccggatc aattccgttt
     6361 tccgctgcat aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt
     6421 ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa
     6481 cggcgtcagc cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc
     6541 actgtccctt attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg
     6601 gctaccgccg gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg
     6661 aagggcagcc cacctatcac ggaattgatc cccctcgaat tggaattc
//
