LOCUS       Exported                7124 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 7124)
  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..7124
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          join(6292..7124,1..6291)
                     /organism="Cloning vector pWW3536"
                     /mol_type="other DNA"
                     /db_xref="taxon:1977344"
     promoter        1..164
                     /label=PBfP2E5
     terminator      2..44
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             169..250
                     /label=RBS1P-LP
     misc_feature    251..853
                     /label=PhlF
     terminator      877..948
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      964..991
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     propeptide      1131..1260
                     /label=pPhlF133
     misc_feature    1182..1211
                     /label=PhlF Operator
     misc_feature    1231..1260
                     /label=PhlF Operator
     RBS             1265..1346
                     /label=RBS1P-LP
     CDS             1347..2060
                     /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             2067..2138
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             2142..2159
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2182..2209
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2441..3724)
                     /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(4164..5024)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71365.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             5512..6246
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71367.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     rep_origin      6312..7085
     rep_origin      6316..6704
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            6875..6984
                     /note="incP origin of transfer"
ORIGIN
        1 gataaaacga aaggctcagt cgaaagactg ggcctttcgt tttacaattg ggctaccttt
       61 tttttgtttt gtttgcaatg gttaatctat tgttaaaatt taaagtttca cttgaacttt
      121 caaataatgt tcttatattt ccagtgtcga aagaaacaaa gtaggactga tcacacaaca
      181 tttaaaaaat aacattatga aagcactcga aaagagattc aaaggtaata ttaacaataa
      241 tttattttca atggcacgta ccccgagccg tagcagcatt ggtagcctgc gtagtccgca
      301 tacccataaa gcaattctga ccagcaccat tgaaatcctg aaagaatgtg gttatagcgg
      361 tctgagcatt gaaagcgttg cacgtcgtgc cggtgcaagc aaaccgacca tttatcgttg
      421 gtggaccaat aaagcagcac tgattgccga agtgtatgaa aatgaaagcg aacaggtgcg
      481 taaatttccg gatctgggta gctttaaagc cgatctggat tttctgctgc gtaatctgtg
      541 gaaagtttgg cgtgaaacca tttgtggtga agcatttcgt tgtgttattg cagaagcaca
      601 gctggaccct gcaaccctga cccagctgaa agatcagttt atggaacgtc gtcgtgagat
      661 gccgaaaaaa ctggttgaaa atgccattag caatggtgaa ctgccgaaag ataccaatcg
      721 tgaactgctg ctggatatga tttttggttt ttgttggtat cgcctgctga ccgaacagct
      781 gaccgttgaa caggatattg aagaatttac cttcctgctg attaatggtg tttgtccggg
      841 tacacagcgt taactagggc ccataccccc aggcatcaaa taaaacgaaa ggctcagtcg
      901 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac
      961 actggctcac cttcgggtgg gcctttctgc gtttatacga cgtacggtgg aatctgattc
     1021 gttaccaatt gacatgatac gaaacgtacc gtatcgacgg tttatacgac gtacggtgga
     1081 atctgattcg ttaccaattg acatgatacg aaacgtaccg tatcgacggc aattgggcta
     1141 cctttttttt gttttgtttg caatggttaa tctattgtta aatgatacga aacgtaccgt
     1201 atcgttaagg tatgttctta tatttgcagt atgatacgaa acgtaccgta tcgttaaggt
     1261 gactgatcac acaacattta aaaaataaca ttatgaaagc actcgaaaag agattcaaag
     1321 gtaatattaa caataattta ttttcaatgc gtaaaggcga agagctgttc actggtgtcg
     1381 tccctattct ggtggaactg gatggtgatg tcaacggtca taagttttcc gtgcgtggcg
     1441 agggtgaagg tgacgcaact aatggtaaac tgacgctgaa gttcatctgt actactggta
     1501 aactgccggt tccttggccg actctggtaa cgacgctgac ttatggtgtt cagtgctttg
     1561 ctcgttatcc ggaccatatg aagcagcatg acttcttcaa gtccgccatg ccggaaggct
     1621 atgtgcagga acgcacgatt tcctttaagg atgacggcac gtacaaaacg cgtgcggaag
     1681 tgaaatttga aggcgatacc ctggtaaacc gcattgagct gaaaggcatt gactttaaag
     1741 aagacggcaa tatcctgggc cataagctgg aatacaattt taacagccac aatgtttaca
     1801 tcaccgccga taaacaaaaa aatggcatta aagcgaattt taaaattcgc cacaacgtgg
     1861 aggatggcag cgtgcagctg gctgatcact accagcaaaa cactccaatc ggtgatggtc
     1921 ctgttctgct gccagacaat cactatctga gcacgcaaag cgttctgtct aaagatccga
     1981 acgagaaacg cgatcatatg gttctgctgg agttcgtaac cgcagcgggc atcacgcatg
     2041 gtatggatga actgtacaaa ggatctgatt acaaggacga tgatgataag gactacaaag
     2101 atgatgatga caaagattat aaggacgatg atgataaagg tcatcatcat catcatcact
     2161 aaatcctagc tgattagctc gagaaggcca tcctgacgga tggccttttt tttgtatcga
     2221 gacgactagt gtaattgcct atcttccagt gatggaacag catttgtgca ttggctgcaa
     2281 caatcagcct tacttgtgcc tgttctattt ccgaaccgac cgcttgtatg aatccatcaa
     2341 aattcgtttt ctctatgttg gattccttgt tgctcatatt gtgatgataa tttctacaaa
     2401 tatagtcatt ggtaactatc tatgaaactg tttgatactt ttatagttga ttaaacttgt
     2461 tcatggcatt tgccttaata tcatccgcta tgtcaatgta gggtttcata gctttgtagt
     2521 cgctgtgtcc cgtccatttc atgaccacct gtgccgggat tccgagagcc agcgcattgc
     2581 agatgaatgt ccttcttcct gcatgggtac tgagcaaagc gtatttgggt gtgacttcat
     2641 caatacgttc atttcccttg tagtaggttt cccgtacagg ctcgttgatt tctgccagtt
     2701 cgcccagctc tttcaggtaa tcgttcatct tctggttgct gatgacgggc agagccatgt
     2761 aattctcgaa atggatgtcc ttgtatttgt ccagtatggc tttgctgtat ttgttcagtt
     2821 caatcgtcag gctgtcggca gtcttgactg tggttatttc gatgtggtcg gacttcacat
     2881 cgcttctttt cagattgcga acatccgaat accgcaaact cgtaaagcag cagaacagga
     2941 aaacatcacg cacacgttcc aggtattgct tatccttggg tatctggtag tctttcagct
     3001 tgttcagttc atcccaagtc aggaagatta cttttttcga ggtggttttc agtttcggtt
     3061 tgaacgtatc gtatgcaatg ttctgatgat gtcctttctt gaagctccag cgcaggaacc
     3121 atttgaggaa tcccatttgc ttgccgatgg tgctgtttct catatccttg gtgtcacgca
     3181 ggaagttgac gtattcgttc aatccaaact cgttgaaata gttgaacgtt gcatcctcct
     3241 tgaactcttt gaggtggttc ctcactgctg caaatttttc ataggtggat gccgtccagt
     3301 tattctggtt accgcactct tttacaaact catcgaacac ctcccaaaag ctgacagggg
     3361 cttcttccgg ctgttcttcg ctggtgtctt tcattctcat gttgaaagct tccttcaact
     3421 gttgggtcgt tggcatgacc tcctgcacct caaattcctt gaaaatattc tggatttcgg
     3481 catagtattt cagcaagtcc gtattgattt cggctgcact ttgctttagc ttgttggtac
     3541 atccgctctt tacccgctgc ttatctgcat cccatttggc tacgtcaatc cggtagcccg
     3601 ttgtaaactc gatgcgttgg ctggcaaaga tgacacgcat acggatgggt acgttctcta
     3661 cgattggcac accgttcttt ttccggctct ccaatgcaaa aatgatgttg cgcttgatat
     3721 tcataattgg gtgcgtttga aattctacac ccaaatatac acccaattat tgagatagca
     3781 aaagacattt agaaacattt acttttactc tatattgtaa tttacacttg attatcagtc
     3841 gtttgcagtc ttatgatatt ctgtgaaagt ataagttcga gagcctgtct ctccgcaaaa
     3901 aacgctgaaa atcagcagat tgcaaaacaa acaccctgtt ttacacccaa gaatgtaaag
     3961 tcggctgttt ttgttttatt taagataata caaccactac ataataaaag agtagcgata
     4021 ttaaaagaat ccgatgagaa aagactaata tttatctatc cattcagttt gatttttcag
     4081 gactttacat cgtcctgaaa gtatttgttg gtaccgagga cgcgacagct gcagacgcgt
     4141 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4201 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4261 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgggaccc acgctcaccg
     4321 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4381 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4441 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4501 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4561 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4621 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4681 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4741 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4801 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4861 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4921 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4981 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5041 tcatggccgc gggattaaaa gtcggggatt ggtgaacaaa aaggtgtttc tctctttaag
     5101 agaaatatcg ttttgctaaa cagttgatat tgaggtatca ttttatcgta aaagacattt
     5161 ttgctcaaca attgcttgac ggaaatcaac aaattttagc attttgtaaa aaagtcgcta
     5221 tataatttgg tgaattggag ttattttcat atttttgcat cccgaagagt ttctcttaaa
     5281 gagagaaaca tcttttgcat accttttccg accgaatttt tatgtcgtaa agaggggctt
     5341 tgcagggggt ggactcagaa agatgagaat agatgactat tgtagttgaa acacatagaa
     5401 agttgctgat atacagaccg atacgcatat cgggatgaac catgagtacg ttcttttctc
     5461 aaaaaacata aatattcgaa aagagatgca ataaattaag gagaggttat aatgaacaaa
     5521 gtaaatataa aagatagtca aaattttatt acttcaaaat atcacataga aaaaataatg
     5581 aattgcataa gtttagatga aaaagataac atctttgaaa taggtgcagg gaaaggtcat
     5641 tttactgctg gattggtaaa gagatgtaat tttgtaacgg cgatagaaat tgattctaaa
     5701 ttatgtgagg taactcgtaa taagctctta aattatccta actatcaaat agtaaatgat
     5761 gatatactga aatttacatt tcctagccac aatccatata aaatatttgg cagcatacct
     5821 tacaacataa gcacaaatat aattcgaaaa attgtttttg aaagttcagc cacaataagt
     5881 tatttaatag tggaatatgg ttttgctaaa atgttattag atacaaacag atcactagca
     5941 ttgctgttaa tggcagaggt agatatttct atattagcaa aaattcctag gtattatttc
     6001 catccaaaac ctaaagtgga tagcacatta attgtattaa aaagaaagcc agcaaaaatg
     6061 gcatttaaag agagaaaaaa atatgaaact tttgtaatga aatgggttaa caaagagtac
     6121 gaaaaactgt ttacaaaaaa tcaatttaat aaagctttaa aacatgcgag aatatatgat
     6181 ataaacaata ttagtttcga acaatttgta tcgctattta atagttataa aatatttaac
     6241 ggctaaaaac aataggccac atgcaactgt aaatgtttac gcgggtaccg acaccgcggt
     6301 ggaggggaat tcccatgtca gccgttaagt gttcctgtgt cactcaaaat tgctttgaga
     6361 ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac cgttaaacct
     6421 taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa accttagagg
     6481 ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct tagtacgtga
     6541 aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg agggtttagt
     6601 tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga aacatgagag
     6661 cttagtacgt actatcaaca ggttgaactg ctgatcttca gatcctctac gccggacgca
     6721 tcgtggccgg atcaattccg ttttccgctg cataaccctg cttcggggtc attatagcga
     6781 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
     6841 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
     6901 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
     6961 atcctgctct gcgaggctgg ccggctaccg ccggcgtaac agatgagggc aagcggatgg
     7021 ctgatgaaac caagccaacc aggaagggca gcccacctat cacggaattg atccccctcg
     7081 aattggaatt cgttcctcga ttcagatcta aaggatccaa tcta
//
