LOCUS       Exported                7892 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 7892)
  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..7892
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1308..1310
                     /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
     RBS             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             69..2372
                     /codon_start=1
                     /product="Striga hermonthica HTL7 strigolactone receptor 
                     with cpGFP inserted after L166 and an LSSmOrange fused to 
                     the C-terminus"
                     /label=rShHTL7cpGFP(L166)
                     /translation="MSSIGLAHNVTILGSGETTVVLGHGYGTDQSVWKLLVPYLVDDYK
                     VLLYDHMGAGTTNPDYFDFDRYSSLEGYSYDLIAILEEFQVSKCIYVGHSMSSMAAAVA
                     SIFRPDLFHKLVMISPTPRLINTEEYYGGFEQKVMDETLRSLDENFKSLSLGTAPLLLA
                     CDASYNVFIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNHYLS
                     VQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEELFTGVVPILV
                     ELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYP
                     DHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDG
                     NILGHKLEYNFNASLESAAMQEYCRTLFNMRPDIACCITRMICGLDLRPYLGHVTVPCH
                     IIQSSNDIMVPVAVGEYLRKNLGGPSVVEVMPTEGHLPHLSMPEVTIPVVLRHIRQDIT
                     DHGEAAAKEAAAKGGMVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYE
                     GFQTVKLKVTKGGPLPFAWDILSPQFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMN
                     FEDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGMEASSERMYPEDGAL
                     KGEDKLRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYER
                     AEGRHSTGGMDELYK"
     primer_bind     564..597
                     /label=D14 GFP insertion_235
     CDS             570..1301
                     /codon_start=1
                     /product="circularly permuted enhanced green fluorescent 
                     protein"
                     /label=cpGFP
                     /translation="YNVFIMADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNFN"
     primer_bind     complement(823..843)
                     /label=D14 GFP insertion_263
     primer_bind     861..878
                     /label=CaMBP SL sensor_303
     primer_bind     complement(1272..1307)
                     /label=D14 GFP insertion_236
     primer_bind     1626..1643
                     /label=D14 GFP insertion_262
     CDS             1665..2372
                     /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     1665..1682
                     /label=CaMBP SL sensor_303
     primer_bind     complement(2353..2408)
                     /label=D14 GFP insertion_263
     terminator      2385..2432
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2607..2635)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     promoter        2748..2852
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2853..3713
                     /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(2880..2905)
                     /label=Marius_81
     rep_origin      3884..4472
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4275..4293
                     /label=Marius_65
     primer_bind     complement(4552..4574)
                     /label=Marius_61
     misc_feature    4658..4800
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4895..4915
                     /label=Marius_66
     CDS             complement(4902..5093)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6405..7487)
                     /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     6475..6492
                     /label=Marius_64
     promoter        complement(7488..7565)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        7874..7892
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatataccat gagtagcatt gggttagcgc acaatgttac gatcctggga agcggagaaa
      121 ctaccgtggt attgggtcac ggatacggca ctgatcagtc tgtgtggaaa ctgcttgtac
      181 cgtacttggt tgatgattac aaggtcttac tttacgatca tatgggggcg ggcacaacca
      241 accctgatta tttcgatttc gaccgctata gttcactgga gggctacagt tacgacttga
      301 tcgcaatttt ggaagagttc caagtatcga agtgcattta cgttggacac tcgatgtctt
      361 cgatggcagc ggcagtcgca tctatttttc gccctgatct gtttcacaaa ttagtaatga
      421 tcagccccac accccgcctt attaacactg aagaatacta cggtggattt gagcaaaagg
      481 ttatggacga gaccctgcgc tcgttagacg aaaactttaa aagcctgtcg ttgggtacgg
      541 caccgttgtt gttggcttgt gatgcatcgt ataacgtctt tatcatggcc gacaagcaga
      601 agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
      661 tcgcctatca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      721 accactacct gagcgtgcag tccaaactga gcaaagaccc caacgagaag cgcgatcaca
      781 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
      841 agggcggtac cggagggagc atggtgagca agggcgagga gctgttcacc ggggtggtgc
      901 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      961 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1021 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     1081 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctaca
     1141 ttcaggagcg caccatcttc ttcaaggacg acggcaacta taagacacgc gctgaggtta
     1201 agttcgaggg cgacactctg gttaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1261 acggcaacat cctgggccat aagcttgaat ataacttcaa cgcgtccttg gagagtgcag
     1321 ccatgcagga gtactgccgt actctgttca acatgcgccc ggacattgct tgctgcatta
     1381 cgcgtatgat ttgcgggctg gatttgcgtc catatctggg acacgtgacc gttccatgcc
     1441 acatcattca aagctcgaac gacatcatgg ttccggtcgc ggtgggggag tacttgcgta
     1501 agaatcttgg tggcccatcg gtcgttgagg tgatgcccac cgagggacat ttaccgcatt
     1561 tgagtatgcc tgaagttact atccctgtag tgttacgtca cattcgccaa gacatcacgg
     1621 accacggaga agcagcagca aaggaagcgg cggcaaaagg gggcatggtg agcaagggcg
     1681 aggagaataa catggccatc atcaaggagt tcatgcgctt caaggtgcgc atggagggct
     1741 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggct
     1801 ttcagaccgt taagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatct
     1861 tgtcccctca gttcacctac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     1921 actacctcaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     1981 acggcggcgt ggtgaccgtg actcaggact cctccctgca ggacggcgag ttcatctaca
     2041 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     2101 tgggcatgga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgagg
     2161 acaagctcag gctgaagctg aaggacggcg gccactacac ctccgaggtc aagaccacct
     2221 acaaggccaa gaagcccgtg cagttgcccg gcgcctacat cgtcgacatc aagttggaca
     2281 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     2341 actccaccgg cggcatggac gagctgtaca aggctgagca ataactagca taaccccttg
     2401 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
     2461 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     2521 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     2581 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     2641 agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2701 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2761 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2821 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2881 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2941 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3001 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3061 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     3121 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3181 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3241 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3301 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3361 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     3421 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3481 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3541 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3601 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3661 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3721 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3781 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3841 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3901 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3961 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4021 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4081 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4141 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4201 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4261 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4321 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     4381 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4441 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4501 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4561 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4621 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     4681 cgcatctgtg cggtatttca caccgcatat atggtgcact ctcagtacaa tctgctctga
     4741 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     4801 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4861 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4921 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     4981 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     5041 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     5101 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     5161 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     5221 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     5281 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     5341 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     5401 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     5461 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     5521 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gccaggaccc aacgctgccc
     5581 gagatgcgcc gcgtgcggct gctggagatg gcggacgcga tggatatgtt ctgccaaggg
     5641 ttggtttgcg cattcacagt tctccgcaag aattgattgg ctccaattct tggagtggtg
     5701 aatccgttag cgaggtgccg ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac
     5761 cgcgacgcaa cgcggggagg cagacaaggt atagggcggc gcctacaatc catgccaacc
     5821 cgttccatgt gctcgccgag gcggcataaa tcgccgtgac gatcagcggt ccagtgatcg
     5881 aagttaggct ggtaagagcc gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta
     5941 cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa
     6001 tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg
     6061 cgtcggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac
     6121 cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga
     6181 tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca
     6241 cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc
     6301 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     6361 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     6421 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6481 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     6541 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     6601 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     6661 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     6721 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     6781 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     6841 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     6901 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     6961 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     7021 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     7081 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     7141 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     7201 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     7261 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     7321 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     7381 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     7441 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     7501 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     7561 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     7621 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     7681 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     7741 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     7801 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc
     7861 gatcccgcga aattaatacg actcactata gg
//
