LOCUS       Exported                7449 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 7449)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 5, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7449
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          7066..7259
                     /organism="Enterobacteria phage lambda"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:10710"
     source          7260..7333
                     /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             1271..1293
                     /label=RBS
                     /note="pLannotate"
                     /note="/database=snapgene"
                     /note="/fragment=False"
                     /note="/identity=100.0"
                     /note="/match_length=100.0"
                     /note="/other=RBS"
     misc_feature    1294..1311
                     /label=TIR med
     CDS             1312..1329
                     /codon_start=1
                     /label=His tag
                     /label=His\tag
                     /note="/vntifkey=31"
                     /translation="HHHHHH"
     CDS             1339..1356
                     /codon_start=1
                     /label=thrombin
                     /note="/vntifkey=31"
                     /translation="LVPRGS"
     CDS             1360..1392
                     /codon_start=1
                     /label=T7 tag
                     /label=T7\tag
                     /note="/vntifkey=31"
                     /translation="MASMTGGQQMG"
     CDS             1396..2109
                     /codon_start=1
                     /product="green fluoresent protein"
                     /label=GFP
                     /note="GFP"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    1407..1415
                     /label=USER
     terminator      2113..2361
                     /label=Tz terminator
     terminator      2125..2167
                     /label=TUUGG terminator
     terminator      2179..2225
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2289..2336
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2372..2400
                     /label=Promoter 3BCD2
                     /note="Promoter 3BCD2"
     misc_feature    2434..2448
                     /label=TIR RK2
     CDS             2440..3630
                     /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      3641..3681
                     /label=rrnC
                     /note="rrnC"
     CDS             complement(3685..4311)
                     /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        4312..4414
                     /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    4432..4441
                     /label=PshAI
                     /note="modified"
     misc_feature    4442..4449
                     /label=SwaI
     misc_feature    4461..4467
                     /label=SanDI
     terminator      4468..4570
                     /label=Lambda T0 terminator
                     /label=T0
     misc_feature    4571..4576
                     /label=speI
     misc_feature    4577..4822
                     /label=OriT
     misc_feature    4823..4830
                     /label=FseI
     rep_origin      4831..7052
                     /label=RK2
     misc_feature    7053..7060
                     /label=AscI
     misc_feature    7066..7333
                     /label=cos
     misc_feature    7248..7259
                     /label=cosN site
     misc_feature    7284..7303
                     /label=ONR455
     misc_feature    7314..7333
                     /label=ONR454
     terminator      complement(7344..7448)
                     /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 aattcccctc
     1261 tagaaataat tttgtttaac tttaagaagg agaatccgga tgggtagcag ccatcatcat
     1321 catcatcaca gcagcggcct ggtgccgcgc ggcagccata tggctagcat gactggtgga
     1381 cagcaaatgg gtcgcatgag caaaggagaa gaacttttca ctggagttgt gccaattctt
     1441 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
     1501 gatgctacat acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt
     1561 ccatggccaa cacttgtcac tactctgacc tatggcgttc aatgcttttc ccgttatccg
     1621 gatcacatga aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa
     1681 cgcactatat ctttcaaaga tgacgggaac tacaagacgc gtgctgaagt caagtttgaa
     1741 ggtgataccc ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac
     1801 attctcggac acaaactaga gtacaactat aactcacaca atgtatacat cacggcagac
     1861 aaacaaaaga atggaatcaa agctaacttc aaaattcgcc acaacattga agatggttcc
     1921 gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta
     1981 ccagacaacc attacctgtc gacacaatct gccctttcga aagatcccaa cgaaaagcgt
     2041 gaccacatgg tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag
     2101 ctctacaaat gatcgaccta gcataacccc gcggggcctc ttcgggggtc tcgcggggtt
     2161 ttttgctgaa agaagcttca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2221 tttatctgtt gtttgtcgcc aaagcccgaa aggaagctga gttggctgct gccaccgctg
     2281 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2341 aaggaggaac tatatccgga taaaaattta tttgcttatt aatcatccgg ctcgtataat
     2401 gtgtggaggg cccaagttca cttaaaaagg agagcgacga tgaaatctaa caatgcgctc
     2461 atcgtcatcc tcggcaccgt caccctggat gctgtaggca taggcttggt tatgccggta
     2521 ctgccgggcc tcttgcggga tatcgtccat tccgacagca tcgccagtca ctatggcgtg
     2581 ctgctagcgc tatatgcgtt gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc
     2641 gaccgctttg gccgccgccc agtcctgctc gcttcgctac ttggagccac tatcgactac
     2701 gcgatcatgg cgaccacacc cgtcctgtgg attctctacg ccggacgcat cgtggccggc
     2761 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     2821 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     2881 cccgtggccg ggggactgtt gggcgccatc tccctgcacg caccattcct tgcggcggcg
     2941 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     3001 gagcgccgtc cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     3061 ggcatgacca ttgtggccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     3121 gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg
     3181 agcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact
     3241 ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac
     3301 gcgctgggct acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg
     3361 attcttctcg cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag
     3421 gtagatgacg accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact
     3481 tcgatcactg gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac
     3541 gggttggcat ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc
     3601 ggtgcatgga gccgggccac ctcgacctga gacaattgtc cagataaaaa aaatccttag
     3661 ctttcgctaa ggatgatttc tttattgtaa ttcattaagc attctgccga catggaagcc
     3721 atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt
     3781 ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa
     3841 atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa
     3901 ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg
     3961 tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg
     4021 ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt
     4081 cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc
     4141 cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg
     4201 aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg
     4261 atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc
     4321 cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt
     4381 atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc agacccttgt
     4441 catttaaatc gtaattattg gggacccctg gattctcacc aataaaaaac gcccggcggc
     4501 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     4561 ggagtccaag actagtcttt tccgctgcat aaccctgctt cggggtcatt atagcgattt
     4621 tttcggtata tccatccttt ttcgcacgat atacaggatt ttgccaaagg gttcgtgtag
     4681 actttccttg gtgtatccaa cggcgtcagc cgggcaggat aggtgaagta ggcccacccg
     4741 cgagcgggtg ttccttcttc actgtccctt attcgcacct ggcggtgctc aacgggaatc
     4801 ctgctctgcg aggctggccg taggccggcc gcgatgcagg tggctgctga acccccagcc
     4861 ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg
     4921 tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac
     4981 ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg
     5041 tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg
     5101 cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc
     5161 tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg
     5221 tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc
     5281 gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag
     5341 cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc
     5401 ttcgcgtact ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg
     5461 ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc
     5521 tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc
     5581 atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg
     5641 atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cttcgctgac ctgttttgcc
     5701 aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc
     5761 atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac gcgaacgctc cacggcggcc
     5821 gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta
     5881 gcttgctgga ctatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg
     5941 cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg
     6001 tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccggaatta
     6061 attccccgga tcgatccgtc gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga
     6121 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     6181 attccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     6241 tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     6301 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgtg gctgccattt
     6361 ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt
     6421 tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg
     6481 cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa
     6541 aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg
     6601 cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc
     6661 tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac
     6721 cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag
     6781 gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc
     6841 ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct
     6901 catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccctacatg
     6961 gctctgctgt agtgagtggg ttgcgctccg gcagcggtcc tgatcccccg cagaaaaaaa
     7021 ggatctcaag aagatccttt gatcttttct acggcgcgcc caataccatt gttcattcca
     7081 cggacaaaaa cagagaaagg aaacgacaga ggccaaaaag cctcgctttc agcacctgtc
     7141 gtttcctttc ttttcagagg gtattttaaa taaaaacatt aagttatgac gaagaagaac
     7201 ggaaacgcct taaaccggaa aattttcata aatagcgaaa acccgcgagg tcgccgcccc
     7261 gtaacctgtc ggatcaccgg aaaggacccg taaagtgata atgattatca tctacatatc
     7321 acaacgtgcg tggattacga ttttctaggg cggcggattt gtcctactca ggagagcgtt
     7381 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     7441 ttgatgcct
//
