LOCUS       Exported                7391 bp ds-DNA     circular SYN 04-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   pSEVA121
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7391)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 5, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7391
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          7009..7201
                     /organism="Enterobacteria phage lambda"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10710"
     source          7202..7275
                     /organism="Enterobacteria phage lambda"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10710"
     misc_feature    1..8
                     /label=PacI
     CDS             complement(18..1100)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(1101..1178)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="lacI promoter"
     misc_feature    1209..1231
                     /label=T7CONS
     promoter        1209..1227
                     /label=T7 promoter
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1232..1256
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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             1268..1294
                     /label=TIR high
                     /note="TIR"
                     /note="high TIR"
     CDS             1286..1341
                     /codon_start=1
                     /label=BCD
                     /translation="MKAIFVLKHLNHAKEVF"
     CDS             1342..2052
                     /codon_start=1
                     /product="green fluoresent protein"
                     /label=GFP
                     /note="GFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      2056..2304
                     /label=Tz terminator
     terminator      2068..2110
                     /label=TUUGG terminator
     terminator      2122..2168
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2232..2279
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2315..2343
                     /label=Promoter 3BCD2
                     /note="Promoter 3BCD2"
     misc_feature    2377..2391
                     /label=TIR RK2
     CDS             2383..3573
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="TcR"
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMSGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     terminator      3584..3624
                     /label=rrnC
                     /note="rrnC"
     CDS             complement(3628..4254)
                     /codon_start=1
                     /label=cat
                     /note="pLannotate"
                     /note="/database=swissprot"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=CDS"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQ"
     promoter        4255..4357
                     /label=cat promoter
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=promoter"
     misc_feature    4375..4380
                     /label=SpeI
     terminator      4381..4483
                     /label=Lambda T0 terminator
                     /label=T0
     misc_feature    4484..4490
                     /label=SanDI
     misc_feature    4491..4501
                     /label=scar1
     misc_feature    4502..4509
                     /label=SwaI
     misc_feature    4510..4519
                     /label=PshAI
                     /note="modified"
     misc_feature    4520..4765
                     /label=OriT
     misc_feature    4766..4773
                     /label=FseI
     rep_origin      4774..6995
                     /label=RK2
     misc_feature    6996..7003
                     /label=AscI
     misc_feature    7009..7275
                     /label=cos
     misc_feature    7190..7201
                     /label=cosN site
     misc_feature    7226..7245
                     /label=ONR455
     misc_feature    7256..7275
                     /label=ONR454
     terminator      complement(7286..7390)
                     /label=T1 terminator
ORIGIN
        1 ttaattaagc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
       61 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
      121 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
      181 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
      241 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
      301 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
      361 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
      421 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
      481 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
      541 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
      601 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
      661 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
      721 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
      781 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
      841 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
      901 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
      961 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     1021 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     1081 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     1141 gccataccgc gaaaggtttt gcgccattcg atggtgtcgg atcgagatcg atctcgatcc
     1201 cgcgaaatta atacgactca ctatagggag aggaattgtg agcggataac aattcctact
     1261 agagattaaa gaggagaaac tatatatgaa agcaattttc gtactgaaac atcttaatca
     1321 tgctaaggag gttttctaat gagcaaagga gaagaacttt tcactggagt tgtgccaatt
     1381 cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa
     1441 ggtgatgcta catacggaaa actcaccctt aaatttattt gcactactgg aaaactacct
     1501 gttccatggc caacacttgt cactactctg acctatggcg ttcaatgctt ttcccgttat
     1561 ccggatcaca tgaaacggca tgactttttc aagagtgcca tgcccgaagg ttatgtacag
     1621 gaacgcacta tatctttcaa agatgacggg aactacaaga cgcgtgctga agtcaagttt
     1681 gaaggtgata cccttgttaa tcgtatcgag ttaaagggta ttgattttaa agaagatgga
     1741 aacattctcg gacacaaact agagtacaac tataactcac acaatgtata catcacggca
     1801 gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacat tgaagatggt
     1861 tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     1921 ttaccagaca accattacct gtcgacacaa tctgcccttt cgaaagatcc caacgaaaag
     1981 cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca tggcatggat
     2041 gagctctaca aatgatcgac ctagcataac cccgcggggc ctcttcgggg gtctcgcggg
     2101 gttttttgct gaaagaagct tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2161 cgttttatct gttgtttgtc gccaaagccc gaaaggaagc tgagttggct gctgccaccg
     2221 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
     2281 tgaaaggagg aactatatcc ggataaaaat ttatttgctt attaatcatc cggctcgtat
     2341 aatgtgtgga gggcccaagt tcacttaaaa aggagagcga cgatgaaatc taacaatgcg
     2401 ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt ggttatgccg
     2461 gtactgccgg gcctcttgcg ggatatcgtc cattccgaca gcatcgccag tcactatggc
     2521 gtgctgctag cgctatatgc gttgatgcaa tttctatgcg cacccgttct cggagcactg
     2581 tccgaccgct ttggccgccg cccagtcctg ctcgcttcgc tacttggagc cactatcgac
     2641 tacgcgatca tggcgaccac acccgtcctg tggattctct acgccggacg catcgtggcc
     2701 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     2761 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     2821 ggccccgtgg ccgggggact gttgggcgcc atctccctgc acgcaccatt ccttgcggcg
     2881 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     2941 ggagagcgcc gtccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     3001 cggggcatga ccattgtggc cgcacttatg actgtcttct ttatcatgca actcgtagga
     3061 caggtgccgg cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg
     3121 atgagcggcc tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc
     3181 actggtcccg ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc
     3241 gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt
     3301 atgattcttc tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg
     3361 caggtagatg acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta
     3421 acttcgatca ctggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg
     3481 aacgggttgg catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt
     3541 cgcggtgcat ggagccgggc cacctcgacc tgagacaatt gtccagataa aaaaaatcct
     3601 tagctttcgc taaggatgat ttctttattg taattcatta agcattctgc cgacatggaa
     3661 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     3721 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     3781 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
     3841 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     3901 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
     3961 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     4021 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     4081 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     4141 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     4201 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     4261 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     4321 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaactagt
     4381 ctggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     4441 ggagttctga ggtcattact ggatctatca acaggagtcc aaggggtccc caataattac
     4501 gatttaaatg acaagggtcc ttttccgctg cataaccctg cttcggggtc attatagcga
     4561 ttttttcggt atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg
     4621 tagactttcc ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac
     4681 ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga
     4741 atcctgctct gcgaggctgg ccgtaggccg gccgcgatgc aggtggctgc tgaaccccca
     4801 gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag
     4861 gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc
     4921 cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc
     4981 gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc
     5041 ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt
     5101 tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag
     5161 cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc
     5221 gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac
     5281 cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg
     5341 cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg
     5401 acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc
     5461 gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt
     5521 ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc
     5581 ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcttcgct gacctgtttt
     5641 gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg
     5701 gtcatcgact tcgccaaacc tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg
     5761 gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc
     5821 gtagcttgct ggactatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg
     5881 gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc
     5941 ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccggaa
     6001 ttaattcccc ggatcgatcc gtcgatcttg atcccctgcg ccatcagatc cttggcggca
     6061 agaaagccat ccagtttact ttgcagggct tcccaacctt accagagggc gccccagctg
     6121 gcaattccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc
     6181 cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     6241 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtggctgcca
     6301 tttttggggt gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg
     6361 cgttagcggg ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac
     6421 gcgcacaggg cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt
     6481 taaaagacag gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt
     6541 ctgcctgtgg acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc
     6601 ccctcaagtg tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa
     6661 taccgcaggg cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat
     6721 caggcgtttt cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc
     6781 tgcccctcat ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc
     6841 cctcatctgt cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccctac
     6901 atggctctgc tgtagtgagt gggttgcgct ccggcagcgg tcctgatccc ccgcagaaaa
     6961 aaaggatctc aagaagatcc tttgatcttt tctacggcgc gcccaatacc attgttcatt
     7021 ccacggacaa aaacagagaa aggaaacgac agaggccaaa aagcctcgct ttcagcacct
     7081 gtcgtttcct ttcttttcag agggtatttt aaataaaaac attaagttat gacgaagaag
     7141 aacggaaacg ccttaaaccg aaaattttca taaatagcga aaacccgcga ggtcgccgcc
     7201 ccgtaacctg tcggatcacc ggaaaggacc cgtaaagtga taatgattat catctacata
     7261 tcacaacgtg cgtggattac gattttctag ggcggcggat ttgtcctact caggagagcg
     7321 ttcaccgaca aacaacagat aaaacgaaag gcccagtctt tcgactgagc ctttcgtttt
     7381 atttgatgcc t
//
