LOCUS       Exported                7397 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 7397)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 5, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7397
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          7015..7207
                     /organism="Enterobacteria phage lambda"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10710"
     source          7208..7281
                     /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"
     promoter        1209..1238
                     /note="Ptrc"
                     /note="The trc promoter combines the -35 region from the
                     trp promoter and the -10 region from the lacUV5 promoter"
     protein_bind    1239..1262
                     /allele="lac operator"
                     /label=lacO
                     /bound_moiety="lac repressor"
                     /note="lac operator"
     RBS             1274..1300
                     /label=TIR - low
                     /note="TIR"
                     /note="low TIR"
     CDS             1292..1347
                     /codon_start=1
                     /label=BCD
                     /translation="MKAIFVLKHLNHAKEVF"
     CDS             1348..2058
                     /codon_start=1
                     /product="green fluoresent protein"
                     /label=GFP
                     /note="GFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      2062..2310
                     /label=Tz terminator
     terminator      2074..2116
                     /label=TUUGG terminator
     terminator      2128..2174
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2238..2285
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2321..2349
                     /label=Promoter 3BCD2
                     /note="Promoter 3BCD2"
     misc_feature    2383..2397
                     /label=TIR RK2
     CDS             2389..3579
                     /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      3590..3630
                     /label=rrnC
                     /note="rrnC"
     CDS             complement(3634..4260)
                     /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        4261..4363
                     /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    4381..4386
                     /label=SpeI
     terminator      4387..4489
                     /label=Lambda T0 terminator
                     /label=T0
     misc_feature    4490..4496
                     /label=SanDI
     misc_feature    4497..4507
                     /label=scar1
     misc_feature    4508..4515
                     /label=SwaI
     misc_feature    4516..4525
                     /label=PshAI
                     /note="modified"
     misc_feature    4526..4771
                     /label=OriT
     misc_feature    4772..4779
                     /label=FseI
     rep_origin      4780..7001
                     /label=RK2
     misc_feature    7002..7009
                     /label=AscI
     misc_feature    7015..7281
                     /label=cos
     misc_feature    7196..7207
                     /label=cosN site
     misc_feature    7232..7251
                     /label=ONR455
     misc_feature    7262..7281
                     /label=ONR454
     terminator      complement(7292..7396)
                     /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 cgcgaaattt gacaattaat catccggctc gtataatgga attgtgagcg gataacaatt
     1261 cctactagag attaaagagg agaaaggggg gatgaaggct attttcgtac tgaaacatct
     1321 taatcatgct aaggaggttt tctaatgagc aaaggagaag aacttttcac tggagttgtg
     1381 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
     1441 ggtgaaggtg atgctacata cggaaaactc acccttaaat ttatttgcac tactggaaaa
     1501 ctacctgttc catggccaac acttgtcact actctgacct atggcgttca atgcttttcc
     1561 cgttatccgg atcacatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
     1621 gtacaggaac gcactatatc tttcaaagat gacgggaact acaagacgcg tgctgaagtc
     1681 aagtttgaag gtgataccct tgttaatcgt atcgagttaa agggtattga ttttaaagaa
     1741 gatggaaaca ttctcggaca caaactagag tacaactata actcacacaa tgtatacatc
     1801 acggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattcgcca caacattgaa
     1861 gatggttccg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     1921 gtccttttac cagacaacca ttacctgtcg acacaatctg ccctttcgaa agatcccaac
     1981 gaaaagcgtg accacatggt ccttcttgag tttgtaactg ctgctgggat tacacatggc
     2041 atggatgagc tctacaaatg atcgacctag cataaccccg cggggcctct tcgggggtct
     2101 cgcggggttt tttgctgaaa gaagcttcaa ataaaacgaa aggctcagtc gaaagactgg
     2161 gcctttcgtt ttatctgttg tttgtcgcca aagcccgaaa ggaagctgag ttggctgctg
     2221 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     2281 ttttgctgaa aggaggaact atatccggat aaaaatttat ttgcttatta atcatccggc
     2341 tcgtataatg tgtggagggc ccaagttcac ttaaaaagga gagcgacgat gaaatctaac
     2401 aatgcgctca tcgtcatcct cggcaccgtc accctggatg ctgtaggcat aggcttggtt
     2461 atgccggtac tgccgggcct cttgcgggat atcgtccatt ccgacagcat cgccagtcac
     2521 tatggcgtgc tgctagcgct atatgcgttg atgcaatttc tatgcgcacc cgttctcgga
     2581 gcactgtccg accgctttgg ccgccgccca gtcctgctcg cttcgctact tggagccact
     2641 atcgactacg cgatcatggc gaccacaccc gtcctgtgga ttctctacgc cggacgcatc
     2701 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     2761 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     2821 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccctgcacgc accattcctt
     2881 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     2941 cataagggag agcgccgtcc gatgcccttg agagccttca acccagtcag ctccttccgg
     3001 tgggcgcggg gcatgaccat tgtggccgca cttatgactg tcttctttat catgcaactc
     3061 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     3121 gcgacgatga gcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     3181 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     3241 gcggccgacg cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc
     3301 cccattatga ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg
     3361 tccaggcagg tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc
     3421 agcctaactt cgatcactgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc
     3481 acatggaacg ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg
     3541 ttgcgtcgcg gtgcatggag ccgggccacc tcgacctgag acaattgtcc agataaaaaa
     3601 aatccttagc tttcgctaag gatgatttct ttattgtaat tcattaagca ttctgccgac
     3661 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     3721 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     3781 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     3841 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     3901 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     3961 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     4021 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     4081 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     4141 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     4201 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     4261 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     4321 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
     4381 actagtctgg attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat
     4441 ccagatggag ttctgaggtc attactggat ctatcaacag gagtccaagg ggtccccaat
     4501 aattacgatt taaatgacaa gggtcctttt ccgctgcata accctgcttc ggggtcatta
     4561 tagcgatttt ttcggtatat ccatcctttt tcgcacgata tacaggattt tgccaaaggg
     4621 ttcgtgtaga ctttccttgg tgtatccaac ggcgtcagcc gggcaggata ggtgaagtag
     4681 gcccacccgc gagcgggtgt tccttcttca ctgtccctta ttcgcacctg gcggtgctca
     4741 acgggaatcc tgctctgcga ggctggccgt aggccggccg cgatgcaggt ggctgctgaa
     4801 cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag gtcatcattg
     4861 acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc gccgacctgc
     4921 tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc
     4981 ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc
     5041 gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg
     5101 acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg gtccaggacg
     5161 cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc
     5221 atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata ccggccattg
     5281 atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata
     5341 ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc gtcggcccgc
     5401 agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat gaccttgttt
     5461 tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga gcgggccgtg
     5521 tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc
     5581 atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc ttcgctgacc
     5641 tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg
     5701 tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc
     5761 acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc
     5821 ttggccgtag cttgctggac tatcgagccg acggactgga aggtttcgcg gggcgcacgc
     5881 atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt
     5941 tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg cgggattgcc
     6001 ccggaattaa ttccccggat cgatccgtcg atcttgatcc cctgcgccat cagatccttg
     6061 gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca gagggcgccc
     6121 cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg
     6181 taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag
     6241 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtgg
     6301 ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga
     6361 ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt cggcgtgcgc
     6421 ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat tggtttaaaa
     6481 gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg
     6541 gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca tctgtcagca
     6601 ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag
     6661 tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg gaaactcgcg
     6721 taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc cggccgaaat
     6781 cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca agtgtcaacg
     6841 tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca acgccggcgg
     6901 ccctacatgg ctctgctgta gtgagtgggt tgcgctccgg cagcggtcct gatcccccgc
     6961 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggcgcgccc aataccattg
     7021 ttcattccac ggacaaaaac agagaaagga aacgacagag gccaaaaagc ctcgctttca
     7081 gcacctgtcg tttcctttct tttcagaggg tattttaaat aaaaacatta agttatgacg
     7141 aagaagaacg gaaacgcctt aaaccgaaaa ttttcataaa tagcgaaaac ccgcgaggtc
     7201 gccgccccgt aacctgtcgg atcaccggaa aggacccgta aagtgataat gattatcatc
     7261 tacatatcac aacgtgcgtg gattacgatt ttctagggcg gcggatttgt cctactcagg
     7321 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     7381 cgttttattt gatgcct
//
