LOCUS       Exported                7403 bp DNA     circular SYN 11-FEB-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pET DuetI Venus9luxA-luxB
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Feb 11, 2023 from SnapGene 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7403
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /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)."
                     /note="color: #31849b"
     RBS             58..63
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
     misc_feature    69..106
                     /label=MCS-1
                     /note="multiple cloning site #1"
                     /note="color: #99ccff"
     CDS             71..73
                     /codon_start=1
                     /product="start codon"
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             113..1195
                     /codon_start=2
                     /gene="luxA"
                     /product="LuxA luciferase subunit"
                     /label=LuxA
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /note="color: #993366"
                     /translation="MKFGNFLLTYQPPQFSQTEVMKRLVKLGRISEECGFDTVWLLEHH
                     FTEFGLLGNPYVAAAYLLGATKKLNVGTAAIVLPTAHPVRQLEDVNLLDQMSKGRFRFG
                     ICRGLYNKDFRVFGTDMNNSRALAECWYGLIKNGMTEGYMEADNEHIKFHKVKVNPAAY
                     SRGGAPVYVVAESASTTEWAAQFGLPMILSWIINTNEKKAQLELYNEVAQEYGHDIHNI
                     DHCLSYITSVDHDSIKAKEICRKFLGHWYDSYVNATTIFDDSDQTRGYDFNKGQWRDFV
                     LKGHKDTNRRIDYSYEINPVGTPQECIDIIQKDIDATGISNICCGFEANGTVDEIIASM
                     KLFQSDVMPFLKEKQRSLLY"
     misc_feature    1198..1215
                     /label=MCS-1
                     /note="multiple cloning site #1"
                     /note="color: #99ccff"
     primer_bind     1236..1255
                     /label=DuetUP2 Primer
                     /note="color: black; sequence: TTGTACACGGCCGCATAATC"
     primer_bind     complement(1236..1255)
                     /label=DuetDOWN1 Primer
                     /note="color: black; sequence: GATTATGCGGCCGTGTACAA"
     promoter        1261..1279
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     1261..1279
                     /label=T7
                     /note="color: black; sequence: TAATACGACTCACTATAGG; added: 
                     2022-12-12"
     protein_bind    1280..1304
                     /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)."
                     /note="color: #31849b"
     RBS             1333..1338
                     /note="ribosome binding site"
                     /note="color: #a6acb3"
     misc_feature    1344..1345
                     /label=MCS-2
                     /note="multiple cloning site #2"
                     /note="color: #99ccff"
     CDS             1347..2330
                     /codon_start=1
                     /gene="luxB"
                     /product="LuxB luciferase subunit"
                     /label=LuxB
                     /note="from the Photorhabdus luminescens bioluminescence 
                     operon"
                     /note="color: #993366"
                     /translation="MKFGLFFLNFINSTTVQEQSIVRMQEITEYVDKLNFEQILVYENH
                     FSDNGVVGAPLTVSGFLLGLTEKIKIGSLNHIITTHHPVRIAEEACLLDQLSEGRFILG
                     FSDCEKKDEMHFFNRPVEYQQQLFEECYEIINDALTTGYCNPDNDFYSFPKISVNPHAY
                     TPGGPRKYVTATSHHIVEWAAKKGIPLIFKWDDSNDVRYEYAERYKAVADKYDVDLSEI
                     DHQLMILVNYNEDSNKAKQETRAFISDYVLEMHPNENFENKLEEIIAENAVGNYTECIT
                     AAKLAIEKCGAKSVLLSFEPMNDLMSQKNVINIVDDNIKKYHMEYT"
     misc_feature    2336..2421
                     /label=MCS-2
                     /note="multiple cloning site #2"
                     /note="color: #99ccff"
     CDS             2349..2393
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /note="color: #cc99b2"
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2431..2449)
                     /label=T7 Terminator Primer
                     /note="color: black; sequence: GCTAGTTATTGCTCAGCGG"
     terminator      2445..2492
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     rep_origin      2529..2984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     CDS             complement(3099..3959)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 3099 .. 3890 / #ccffcc
                        2: 3891 .. 3959 / #ccffcc / signal sequence
                     Cleavage site after base 3890"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3960..4052)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      4133..4721
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(5151..5342)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /note="color: #993366"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5914..6996)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6997..7074)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     7341..7356
                     /label=pET Upstream Primer
                     /note="color: black; sequence: ATGCGTCCGGCGTAGA"
     promoter        join(7387..7403,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     join(7387..7403,1..2)
                     /label=T7
                     /note="color: black; sequence: TAATACGACTCACTATAGG; added: 
                     2022-12-12"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgatgaaatt
      121 tggaaacttt ttgcttacat accaacctcc ccaattttct caaacagagg taatgaaacg
      181 tttggttaaa ttaggtcgca tctctgagga gtgtggtttt gataccgtat ggttactgga
      241 gcatcatttc acggagtttg gtttgcttgg taacccttat gtcgctgctg catatttact
      301 tggcgcgact aaaaaattga atgtaggaac tgccgctatt gttcttccca cagcccatcc
      361 agtacgccaa cttgaagatg tgaatttatt ggatcaaatg tcaaaaggac gatttcggtt
      421 tggtatttgc cgagggcttt acaacaagga ctttcgcgta ttcggcacag atatgaataa
      481 cagtcgcgcc ttagcggaat gctggtacgg gctgataaag aatggcatga cagagggata
      541 tatggaagct gataatgaac atatcaagtt ccataaggta aaagtaaacc ccgcggcgta
      601 tagcagaggt ggcgcaccgg tttatgtggt ggctgaatca gcttcgacga ctgagtgggc
      661 tgctcaattt ggcctaccga tgatattaag ttggattata aatactaacg aaaagaaagc
      721 acaacttgag ctttataatg aagtggctca agaatatggg cacgatattc ataatatcga
      781 ccattgctta tcatatataa catctgtaga tcatgactca attaaagcga aagagatttg
      841 ccggaaattt ctggggcatt ggtatgattc ttatgtgaat gctacgacta tttttgatga
      901 ttcagaccaa acaagaggtt atgatttcaa taaagggcag tggcgtgact ttgtattaaa
      961 aggacataaa gatactaatc gccgtattga ttacagttac gaaatcaatc ccgtgggaac
     1021 gccgcaggaa tgtattgaca taattcaaaa agacattgat gctacaggaa tatcaaatat
     1081 ttgttgtgga tttgaagcta atggaacagt agacgaaatt attgcttcca tgaagctctt
     1141 ccagtctgat gtcatgccat ttcttaaaga aaaacaacgt tcgctattat attaggcggc
     1201 cgcataatgc ttaagtcgaa cagaaagtaa tcgtattgta cacggccgca taatcgaaat
     1261 taatacgact cactataggg gaattgtgag cggataacaa ttccccatct tagtatatta
     1321 gttaagtata agaaggagat atacatatga aatttggatt gttcttcctt aacttcatca
     1381 attcaacaac tgttcaagaa caaagtatag ttcgcatgca ggaaataacg gagtatgttg
     1441 ataagttgaa ttttgaacag attttagtgt atgaaaatca tttttcagat aatggtgttg
     1501 tcggcgctcc tctgactgtt tctggttttc tgctcggttt aacagagaaa attaaaattg
     1561 gttcattaaa tcacatcatt acaactcatc atcctgtccg catagcggag gaagcttgct
     1621 tattggatca gttaagtgaa gggagattta ttttagggtt tagtgattgc gaaaaaaaag
     1681 atgaaatgca tttttttaat cgcccggttg aatatcaaca gcaactattt gaagagtgtt
     1741 atgaaatcat taacgatgct ttaacaacag gctattgtaa tccagataac gatttttata
     1801 gcttccctaa aatatctgta aatccccatg cttatacgcc aggcggacct cggaaatatg
     1861 taacagcaac cagtcatcat attgttgagt gggcggccaa aaaaggtatt cctctcatct
     1921 ttaagtggga tgattctaat gatgttagat atgaatatgc tgaaagatat aaagccgttg
     1981 cggataaata tgacgttgac ctatcagaga tagaccatca gttaatgata ttagttaact
     2041 ataacgaaga tagtaataaa gctaaacaag agacgcgtgc atttattagt gattatgttc
     2101 ttgaaatgca ccctaatgaa aatttcgaaa ataaacttga agaaataatt gcagaaaacg
     2161 ctgtcggaaa ttatacggag tgtataactg cggctaagtt ggcaattgaa aagtgtggtg
     2221 cgaaaagtgt attgctgtcc tttgaaccaa tgaatgattt gatgagccaa aaaaatgtaa
     2281 tcaatattgt tgatgataat attaagaagt accacatgga atatacctaa ggtaccctcg
     2341 agtctggtaa agaaaccgct gctgcgaaat ttgaacgcca gcacatggac tcgtctacta
     2401 gcgcagctta attaacctag gctgctgcca ccgctgagca ataactagca taaccccttg
     2461 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
     2521 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2581 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2641 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2701 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2761 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2821 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2881 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2941 aaaatttaac gcgaatttta acaaaatatt aacgtttaca atttctggcg gcacgatggc
     3001 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     3061 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     3121 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3181 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     3241 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     3301 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     3361 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     3421 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     3481 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     3541 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     3601 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     3661 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     3721 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3781 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3841 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3901 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3961 ctcttccttt ttcaatcatg attgaagcat ttatcagggt tattgtctca tgagcggata
     4021 catatttgaa tgtatttaga aaaataaaca aataggtcat gaccaaaatc ccttaacgtg
     4081 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4141 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4201 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4261 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4321 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4381 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4441 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4501 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4561 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4621 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4681 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4741 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4801 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     4861 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     4921 ttctccttac gcatctgtgc ggtatttcac accgcatata tggtgcactc tcagtacaat
     4981 ctgctctgat gccgcatagt taagccagta tacactccgc tatcgctacg tgactgggtc
     5041 atggctgcgc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5101 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5161 tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga
     5221 agcgattcac agatgtctgc ctgttcatcc gcgtccagct cgttgagttt ctccagaagc
     5281 gttaatgtct ggcttctgat aaagcgggcc atgttaaggg cggttttttc ctgtttggtc
     5341 actgatgcct ccgtgtaagg gggatttctg ttcatggggg taatgatacc gatgaaacga
     5401 gagaggatgc tcacgatacg ggttactgat gatgaacatg cccggttact ggaacgttgt
     5461 gagggtaaac aactggcggt atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa
     5521 tgccagcgct tcgttaatac agatgtaggt gttccacagg gtagccagca gcatcctgcg
     5581 atgcagatcc ggaacataat ggtgcagggc gctgacttcc gcgtttccag actttacgaa
     5641 acacggaaac cgaagaccat tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag
     5701 tcgcttcacg ttcgctcgcg tatcggtgat tcattctgct aaccagtaag gcaaccccgc
     5761 cagcctagcc gggtcctcaa cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac
     5821 cggaaggagc tgactgggtt gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa
     5881 tgagtgagct aacttacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5941 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6001 gggcgccagg gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac
     6061 cgcctggccc tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa
     6121 atcctgtttg atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta
     6181 tcccactacc gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc
     6241 gcccagcgcc atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag
     6301 catttgcatg gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat
     6361 cggctgaatt tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga
     6421 gacagaactt aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg
     6481 ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg
     6541 gtcagagaca tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc
     6601 atcctggtca tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt
     6661 gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct
     6721 ggcacccagt tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag
     6781 ggccagactg gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc
     6841 cacgcggttg ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt
     6901 cgcagaaacg tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc
     6961 atactctgcg acatcgtata acgttactgg tttcacattc accaccctga attgactctc
     7021 ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat
     7081 ctcgacgctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     7141 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     7201 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     7261 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     7321 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg
     7381 cgaaattaat acgactcact ata
//
