LOCUS       Exported                7570 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 7570)
  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..7570
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          join(6738..7570,1..6737)
                     /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..1330
                     /label=EM_gBlock2
     CDS             251..1330
                     /codon_start=1
                     /label=LacI
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLE"
     misc_feature    1425..1551
                     /label=EM_gBlock2
     terminator      1440..1526
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        1587..1706
                     /label=pLacI43
     protein_bind    1658..1674
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1711..1792
                     /label=RBS1P-LP
     CDS             1793..2506
                     /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             2513..2584
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             2588..2605
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2628..2655
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(2887..4170)
                     /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(4610..5470)
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /protein_id="ARI71365.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             5958..6692
                     /codon_start=1
                     /transl_table=11
                     /product="ErmG"
                     /label=ErmG
                     /protein_id="ARI71367.1"
                     /translation="MNKVNIKDSQNFITSKYHIEKIMNCISLDEKDNIFEIGAGKGHFT
                     AGLVKRCNFVTAIEIDSKLCEVTRNKLLNYPNYQIVNDDILKFTFPSHNPYKIFGSIPY
                     NISTNIIRKIVFESSATISYLIVEYGFAKMLLDTNRSLALLLMAEVDISILAKIPRYYF
                     HPKPKVDSTLIVLKRKPAKMAFKERKKYETFVMKWVNKEYEKLFTKNQFNKALKHARIY
                     DINNISFEQFVSLFNSYKIFNG"
     rep_origin      6758..7531
     rep_origin      6762..7150
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     oriT            7321..7430
                     /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 atggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
      301 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
      361 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
      421 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
      481 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
      541 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
      601 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
      661 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
      721 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
      781 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
      841 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
      901 ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa
      961 tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat
     1021 gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag tgggatacga
     1081 cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg
     1141 cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa
     1201 gggcaatcag ctgttgccag tttcactggt gaaaagaaaa accaccctgg cgcccaatac
     1261 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     1321 ccgactggaa gagacgtaag tggagtgacc ttaaatcaag gacgatatta atcggaagga
     1381 gtattcaacg tgatgaagtc gcagggttga cgtgggacgt ctctgatcgg atctccaggc
     1441 atataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1501 tgaacgctct cctgagtagg acaaatccca aagcactgct cgctttgtta agacggccca
     1561 aagcactgct cgctttgtta agacggcaat tgggctacct tttttttgtt ttgtttgcaa
     1621 tggttaatct attgttaaaa tttaaagttt cacttgattg tgagcggata acaattatat
     1681 ttgcagtgtc gaaagaaaca aagtaggact gatcacacaa catttaaaaa ataacattat
     1741 gaaagcactc gaaaagagat tcaaaggtaa tattaacaat aatttatttt caatgcgtaa
     1801 aggcgaagag ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa
     1861 cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac
     1921 gctgaagttc atctgtacta ctggtaaact gccggttcct tggccgactc tggtaacgac
     1981 gctgacttat ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt
     2041 cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga
     2101 cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat
     2161 tgagctgaaa ggcattgact ttaaagaaga cggcaatatc ctgggccata agctggaata
     2221 caattttaac agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc
     2281 gaattttaaa attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca
     2341 gcaaaacact ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac
     2401 gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt
     2461 cgtaaccgca gcgggcatca cgcatggtat ggatgaactg tacaaaggat ctgattacaa
     2521 ggacgatgat gataaggact acaaagatga tgatgacaaa gattataagg acgatgatga
     2581 taaaggtcat catcatcatc atcactaaat cctagctgat tagctcgaga aggccatcct
     2641 gacggatggc cttttttttg tatcgagacg actagtgtaa ttgcctatct tccagtgatg
     2701 gaacagcatt tgtgcattgg ctgcaacaat cagccttact tgtgcctgtt ctatttccga
     2761 accgaccgct tgtatgaatc catcaaaatt cgttttctct atgttggatt ccttgttgct
     2821 catattgtga tgataatttc tacaaatata gtcattggta actatctatg aaactgtttg
     2881 atacttttat agttgattaa acttgttcat ggcatttgcc ttaatatcat ccgctatgtc
     2941 aatgtagggt ttcatagctt tgtagtcgct gtgtcccgtc catttcatga ccacctgtgc
     3001 cgggattccg agagccagcg cattgcagat gaatgtcctt cttcctgcat gggtactgag
     3061 caaagcgtat ttgggtgtga cttcatcaat acgttcattt cccttgtagt aggtttcccg
     3121 tacaggctcg ttgatttctg ccagttcgcc cagctctttc aggtaatcgt tcatcttctg
     3181 gttgctgatg acgggcagag ccatgtaatt ctcgaaatgg atgtccttgt atttgtccag
     3241 tatggctttg ctgtatttgt tcagttcaat cgtcaggctg tcggcagtct tgactgtggt
     3301 tatttcgatg tggtcggact tcacatcgct tcttttcaga ttgcgaacat ccgaataccg
     3361 caaactcgta aagcagcaga acaggaaaac atcacgcaca cgttccaggt attgcttatc
     3421 cttgggtatc tggtagtctt tcagcttgtt cagttcatcc caagtcagga agattacttt
     3481 tttcgaggtg gttttcagtt tcggtttgaa cgtatcgtat gcaatgttct gatgatgtcc
     3541 tttcttgaag ctccagcgca ggaaccattt gaggaatccc atttgcttgc cgatggtgct
     3601 gtttctcata tccttggtgt cacgcaggaa gttgacgtat tcgttcaatc caaactcgtt
     3661 gaaatagttg aacgttgcat cctccttgaa ctctttgagg tggttcctca ctgctgcaaa
     3721 tttttcatag gtggatgccg tccagttatt ctggttaccg cactctttta caaactcatc
     3781 gaacacctcc caaaagctga caggggcttc ttccggctgt tcttcgctgg tgtctttcat
     3841 tctcatgttg aaagcttcct tcaactgttg ggtcgttggc atgacctcct gcacctcaaa
     3901 ttccttgaaa atattctgga tttcggcata gtatttcagc aagtccgtat tgatttcggc
     3961 tgcactttgc tttagcttgt tggtacatcc gctctttacc cgctgcttat ctgcatccca
     4021 tttggctacg tcaatccggt agcccgttgt aaactcgatg cgttggctgg caaagatgac
     4081 acgcatacgg atgggtacgt tctctacgat tggcacaccg ttctttttcc ggctctccaa
     4141 tgcaaaaatg atgttgcgct tgatattcat aattgggtgc gtttgaaatt ctacacccaa
     4201 atatacaccc aattattgag atagcaaaag acatttagaa acatttactt ttactctata
     4261 ttgtaattta cacttgatta tcagtcgttt gcagtcttat gatattctgt gaaagtataa
     4321 gttcgagagc ctgtctctcc gcaaaaaacg ctgaaaatca gcagattgca aaacaaacac
     4381 cctgttttac acccaagaat gtaaagtcgg ctgtttttgt tttatttaag ataatacaac
     4441 cactacataa taaaagagta gcgatattaa aagaatccga tgagaaaaga ctaatattta
     4501 tctatccatt cagtttgatt tttcaggact ttacatcgtc ctgaaagtat ttgttggtac
     4561 cgaggacgcg acagctgcag acgcgttatg agtaaacttg gtctgacagt taccaatgct
     4621 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4681 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4741 tgataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4801 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4861 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4921 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4981 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5041 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5101 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5161 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5221 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5281 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5341 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5401 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5461 gaatactcat actcttcctt tttcaatcat ggccgcggga ttaaaagtcg gggattggtg
     5521 aacaaaaagg tgtttctctc tttaagagaa atatcgtttt gctaaacagt tgatattgag
     5581 gtatcatttt atcgtaaaag acatttttgc tcaacaattg cttgacggaa atcaacaaat
     5641 tttagcattt tgtaaaaaag tcgctatata atttggtgaa ttggagttat tttcatattt
     5701 ttgcatcccg aagagtttct cttaaagaga gaaacatctt ttgcatacct tttccgaccg
     5761 aatttttatg tcgtaaagag gggctttgca gggggtggac tcagaaagat gagaatagat
     5821 gactattgta gttgaaacac atagaaagtt gctgatatac agaccgatac gcatatcggg
     5881 atgaaccatg agtacgttct tttctcaaaa aacataaata ttcgaaaaga gatgcaataa
     5941 attaaggaga ggttataatg aacaaagtaa atataaaaga tagtcaaaat tttattactt
     6001 caaaatatca catagaaaaa ataatgaatt gcataagttt agatgaaaaa gataacatct
     6061 ttgaaatagg tgcagggaaa ggtcatttta ctgctggatt ggtaaagaga tgtaattttg
     6121 taacggcgat agaaattgat tctaaattat gtgaggtaac tcgtaataag ctcttaaatt
     6181 atcctaacta tcaaatagta aatgatgata tactgaaatt tacatttcct agccacaatc
     6241 catataaaat atttggcagc ataccttaca acataagcac aaatataatt cgaaaaattg
     6301 tttttgaaag ttcagccaca ataagttatt taatagtgga atatggtttt gctaaaatgt
     6361 tattagatac aaacagatca ctagcattgc tgttaatggc agaggtagat atttctatat
     6421 tagcaaaaat tcctaggtat tatttccatc caaaacctaa agtggatagc acattaattg
     6481 tattaaaaag aaagccagca aaaatggcat ttaaagagag aaaaaaatat gaaacttttg
     6541 taatgaaatg ggttaacaaa gagtacgaaa aactgtttac aaaaaatcaa tttaataaag
     6601 ctttaaaaca tgcgagaata tatgatataa acaatattag tttcgaacaa tttgtatcgc
     6661 tatttaatag ttataaaata tttaacggct aaaaacaata ggccacatgc aactgtaaat
     6721 gtttacgcgg gtaccgacac cgcggtggag gggaattccc atgtcagccg ttaagtgttc
     6781 ctgtgtcact caaaattgct ttgagaggct ctaagggctt ctcagtgcgt tacatccctg
     6841 gcttgttgtc cacaaccgtt aaaccttaaa agctttaaaa gccttatata ttcttttttt
     6901 tcttataaaa cttaaaacct tagaggctat ttaagttgct gatttatatt aattttattg
     6961 ttcaaacatg agagcttagt acgtgaaaca tgagagctta gtacgttagc catgagagct
     7021 tagtacgtta gccatgaggg tttagttcgt taaacatgag agcttagtac gttaaacatg
     7081 agagcttagt acgtgaaaca tgagagctta gtacgtacta tcaacaggtt gaactgctga
     7141 tcttcagatc ctctacgccg gacgcatcgt ggccggatca attccgtttt ccgctgcata
     7201 accctgcttc ggggtcatta tagcgatttt ttcggtatat ccatcctttt tcgcacgata
     7261 tacaggattt tgccaaaggg ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc
     7321 gggcaggata ggtgaagtag gcccacccgc gagcgggtgt tccttcttca ctgtccctta
     7381 ttcgcacctg gcggtgctca acgggaatcc tgctctgcga ggctggccgg ctaccgccgg
     7441 cgtaacagat gagggcaagc ggatggctga tgaaaccaag ccaaccagga agggcagccc
     7501 acctatcacg gaattgatcc ccctcgaatt ggaattcgtt cctcgattca gatctaaagg
     7561 atccaatcta
//
