LOCUS       Exported                7880 bp ds-DNA     circular SYN 17-JAN-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7880)
  AUTHORS   Rebecca Wood
  TITLE     Direct Submission
  JOURNAL   Exported Jan 17, 2021 from SnapGene 4.1.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7880
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          561..608
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    1..25
                     /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)."
     primer_bind     34..59
                     /label=E coli CL operon_226
     primer_bind     38..90
                     /label=D14 GFP insertion_253
     RBS             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             69..2360
                     /codon_start=1
                     /product="Petunia hybrida DAD2 strigolactone receptor with 
                     cpGFP inserted after G164 and an LSSmOrange fused to the 
                     C-terminus"
                     /label=rDAD2cpGFP(G164)
                     /label=rDAD2cpGFP(G166)
                     /translation="MGQTLLDALNVRVVGSGERVLVLAHGFGTDQSAWNRILPFFLRDY
                     RVVLYDLVCAGSVNPDFFDFRRYTTLDPYVDDLLHILDALGIDCCAYVGHSVSAMIGIL
                     ASIRRPELFSKLILIGASPRFLNDEDYHGGFEQGEIEKVFSAMEANYEAWVNGFAPLAV
                     GADVPAAVREFSRTLFNASYNVFIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTP
                     IGDGPVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMV
                     SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTL
                     VTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTL
                     VNRIELKGIDFKEDGNILGHKLEYNFNASMRPDITLFVSRTVFNSDMRGVLGLVKVPCH
                     IFQTARDHSVPASVATYLKNHLGGKNTVHWLNIEGHLPHLSAPTLLAQELRRALSHRGE
                     AAAKEAAAKGGMVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGFQT
                     VKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDG
                     GVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALKGED
                     KLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERAEGR
                     HSTGGMDELYK"
     misc_feature    175
                     /label=nucleotide called as C, equally could be G
     misc_feature    356
                     /label=nucleotide called as C, definitely appears to be A
     primer_bind     complement(543..638)
                     /label=D14 GFP insertion_242
     primer_bind     561..578
                     /label=D14 GFP insertion_241
     primer_bind     609..642
                     /label=D14 GFP insertion_235
     CDS             615..1346
                     /codon_start=1
                     /product="circularly permuted enhanced green fluorescent 
                     protein"
                     /label=cpGFP
                     /translation="YNVFIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNFN"
     primer_bind     complement(868..888)
                     /label=D14 GFP insertion_263
     primer_bind     906..923
                     /label=CaMBP SL sensor_303
     primer_bind     complement(1317..1352)
                     /label=D14 GFP insertion_236
     primer_bind     1323..1352
                     /label=D14 GFP insertion_241
     primer_bind     1584..1631
                     /label=D14 GFP insertion_262
     primer_bind     complement(1596..1643)
                     /label=D14 GFP insertion_261
     CDS             1653..2360
                     /codon_start=1
                     /product="monomeric orange fluorescent protein with a large
                     Stokes shift (Shcherbakova et al., 2012)"
                     /label=LSSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGALK
                     GEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     1653..1670
                     /label=CaMBP SL sensor_303
     primer_bind     complement(2341..2396)
                     /label=D14 GFP insertion_263
     terminator      2373..2420
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2595..2623)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2736..2840
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2841..3701
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2868..2893)
                     /label=Marius_81
     rep_origin      3872..4460
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4263..4281
                     /label=Marius_65
     primer_bind     complement(4540..4562)
                     /label=Marius_61
     misc_feature    4646..4788
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4883..4903
                     /label=Marius_66
     CDS             complement(4890..5081)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6393..7475)
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6463..6480
                     /label=Marius_64
     promoter        complement(7476..7553)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        7862..7880
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatataccat gggtcaaact ttactggacg ccttgaatgt gagggttgta ggcagcggag
      121 agagagtgtt ggtacttgct cacgggtttg gtaccgacca gtccgcttgg aataggattc
      181 tacccttttt ccttagggac tatagagttg tactatacga tttggtgtgt gcaggatcag
      241 ttaaccctga cttctttgat tttaggaggt acacaacact tgacccttat gtggatgatt
      301 tgctgcatat attagacgcg ctggggatag actgttgcgc ttatgttggt cattcagtat
      361 cagcaatgat cggcatattg gcgtctatac gtaggccgga attattttct aagctaattt
      421 tgataggcgc gagtcctagg tttctaaacg atgaagacta ccacggcggt ttcgagcaag
      481 gagaaatcga aaaggtcttt tccgcgatgg aggcaaacta tgaggcgtgg gtgaatggat
      541 tcgcgccttt ggcagtgggc gctgatgtac cggcagcggt aagggaattc tcccgtacac
      601 tgtttaacgc atcgtataac gtctttatca tggccgacaa gcagaagaac ggcatcaagg
      661 cgaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc tatcactacc
      721 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg
      781 tgcagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      841 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggc ggtaccggag
      901 ggagcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      961 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     1021 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     1081 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     1141 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacattcag gagcgcacca
     1201 tcttcttcaa ggacgacggc aactataaga cacgcgctga ggttaagttc gagggcgaca
     1261 ctctggttaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1321 gccataagct tgaatataac ttcaacgcgt ccatgcgtcc agacataact ttgttcgtta
     1381 gtaggacagt attcaacagc gatatgagag gtgttttggg cttagtgaag gtcccttgtc
     1441 atatctttca gacagctaga gatcattctg tgcctgcgag tgtagccact tatctaaaga
     1501 accacctagg cggtaaaaat acagtgcact ggcttaacat agaaggtcac cttccacatc
     1561 taagtgcgcc cactctttta gcacaagagc ttaggcgtgc actgtctcat cgtggagaag
     1621 cagcagcaaa ggaagcggcg gcaaaagggg gcatggtgag caagggcgag gagaataaca
     1681 tggccatcat caaggagttc atgcgcttca aggtgcgcat ggagggctcc gtgaacggcc
     1741 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcttt cagaccgtta
     1801 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcttg tcccctcagt
     1861 tcacctacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacctcaagc
     1921 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg
     1981 tgaccgtgac tcaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     2041 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg ggcatggagg
     2101 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgaggac aagctcaggc
     2161 tgaagctgaa ggacggcggc cactacacct ccgaggtcaa gaccacctac aaggccaaga
     2221 agcccgtgca gttgcccggc gcctacatcg tcgacatcaa gttggacatc acctcccaca
     2281 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     2341 gcatggacga gctgtacaag gctgagcaat aactagcata accccttggg gcctctaaac
     2401 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
     2461 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
     2521 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
     2581 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
     2641 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     2701 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     2761 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     2821 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     2881 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     2941 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3001 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     3061 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     3121 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     3181 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     3241 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     3301 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     3361 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     3421 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     3481 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     3541 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     3601 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     3661 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     3721 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     3781 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     3841 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     3901 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     3961 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     4021 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     4081 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     4141 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     4201 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     4261 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     4321 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     4381 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     4441 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     4501 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     4561 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     4621 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     4681 gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4741 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     4801 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4861 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4921 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     4981 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     5041 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     5101 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     5161 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     5221 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     5281 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     5341 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     5401 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     5461 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     5521 gacaggagca cgatcatgcg cacccgtggc caggacccaa cgctgcccga gatgcgccgc
     5581 gtgcggctgc tggagatggc ggacgcgatg gatatgttct gccaagggtt ggtttgcgca
     5641 ttcacagttc tccgcaagaa ttgattggct ccaattcttg gagtggtgaa tccgttagcg
     5701 aggtgccgcc ggcttccatt caggtcgagg tggcccggct ccatgcaccg cgacgcaacg
     5761 cggggaggca gacaaggtat agggcggcgc ctacaatcca tgccaacccg ttccatgtgc
     5821 tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc agtgatcgaa gttaggctgg
     5881 taagagccgc gagcgatcct tgaagctgtc cctgatggtc gtcatctacc tgcctggaca
     5941 gcatggcctg caacgcgggc atcccgatgc cgccggaagc gagaagaatc ataatgggga
     6001 aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca
     6061 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     6121 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     6181 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     6241 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     6301 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     6361 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6421 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6481 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     6541 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     6601 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     6661 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     6721 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     6781 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     6841 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     6901 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     6961 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     7021 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     7081 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     7141 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     7201 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     7261 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     7321 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     7381 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     7441 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     7501 tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc
     7561 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     7621 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     7681 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     7741 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     7801 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
     7861 ttaatacgac tcactatagg
//
