LOCUS       Exported                5683 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 5683)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 5, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5683
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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)."
     primer_bind     complement(1233..1326)
                     /label=7580 His_on_SAGA_rev
     primer_bind     complement(1268..1323)
                     /label=Tags_Rand_rev2
     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=2
                     /product="green fluoresent protein"
                     /label=GFP
                     /note="GFP"
                     /translation="*AKEKNFSLELCQFLLN*MVMLMGTNFLSVERVKVMLHTENSPLN
                     LFALLENYLFHGQHLSLL*PMAFNAFPVIRIT*NGMTFSRVPCPKVMYRNALYLSKMTG
                     TTRRVLKSSLKVIPLLIVSS*RVLILKKMETFSDTN*STTITHTMYTSRQTNKRMESKL
                     TSKFATTLKMVPFN*QTIINKILQLAMALSFYQTTITCRHNLPFRKIPTKSVTTWSFLS
                     L*LLLGLHMAWMSST"
     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_medium
     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..4437
                     /label=SpeI
     terminator      4438..4540
                     /label=Lambda T0 terminator
                     /label=T0
     misc_feature    4541..4547
                     /label=SanDI
     misc_feature    4559..4566
                     /label=SwaI
     misc_feature    4567..4576
                     /label=PshAI
                     /note="modified"
     misc_feature    4577..4822
                     /label=OriT
     misc_feature    4823..4830
                     /label=FseI
     rep_origin      4867..5412
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     misc_feature    5564..5571
                     /label=AscI
     terminator      5572..5683
                     /label=T1
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 agaccaggga tgaagtccaa 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 aactagtctg
     4441 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     4501 gttctgaggt cattactgga tctatcaaca ggagtccaag gggtccccaa taattacgat
     4561 ttaaatgaca agggtccttt 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 ctagaaatat tttatctgat taataagatg
     4861 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     4921 cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg
     4981 gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga
     5041 cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat
     5101 gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa
     5161 cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca
     5221 ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca
     5281 ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct
     5341 gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg
     5401 agcctatgga aaaacggctt ttgccgcggc cctctcactt ccctgttaag tatcttcctg
     5461 gcatcttcca ggaaatctcc gccccgttcg taagccattt ccgctcgccg cagtcgaacg
     5521 accgagcgta gcgagtcagt gagcgaggaa gcggaatata tccggcgcgc ccagctgtct
     5581 agggcggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     5641 aggcccagtc tttcgactga gcctttcgtt ttatttgatg cct
//
