LOCUS       Exported                7857 bp ds-DNA     circular SYN 11-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7857)
  AUTHORS   Hehnly Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Oct 9, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7857
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     misc_feature    101..617
                     /label=CIB1
     primer_bind     591..649
                     /label=Ffrag-cib1-rfp-ift88a
     regulatory      629..638
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             635..1366
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     misc_feature    1368..1384
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1391..3868
                     /codon_start=1
                     /label=IFT88
                     /translation="MMENVHLAPETDEDDLYSGFNDYNPAYDTEELENDTGFQQAVRTS
                     HGRRPPVTAKIPSTAVSRPIATGYGSKTSLTSSMGRPMTGTIQDGVARPMTAVRAAGFS
                     KAALRGSAFDPLGQSRGPAPPLEAKNEDSPEEKIRQLEKKVNELVEESCIANSCGDLKL
                     ALEKAKDAGRKERVLVRQREQVTSPENINLDLTYSVLFNLASQYSANEMYAEALNTYQV
                     IVKNKMFSNAGRLKVNMGNIYLRQRNYSKAIKFYRMALDQIPSVHKEMRIKIMQNIGIT
                     FIKTGQYSDAINSFEHIMSMAPSLKAGFNLILSCFAIGDREKMKKAFQKLIAVPLEIDE
                     DDKYISPSDDPHTNLLIEAIKNDHLRQMERERKAMAEKYIMTAAKLIAPVIEASFAVGY
                     NWCVEVVKASQYVELANDLEINKAITYLRQKDFNQAVDTLKMFERKDSRVKSAAATNLS
                     FLYYLENEFAQASSYADLAVNSDRYNPSALTNKGDTVFANGDYEKAAEFYKEALRNDSS
                     CTEALYNIGLTYKKLNRLDEALDSFLKLHAILRNSAQVLCQIANIYELMEDPNQAIEWL
                     MQLISVVPTDSQALSKLGELYDSEGDKSQAFQYYYESYRYFPSNIEVIEWLGAYYIDTQ
                     FCEKAIQYFERASLIQPTQVKWQLMVASCFRRSGNYQKALDTYKEIHRKFPGNVECLRF
                     LVRLCTDIGLKEVQEYATKLKRLEKMKEMREQRIKSGRDSSGGSRSKREGSAGSDSGQN
                     NSASSKSERLSAKLRALPGTDEPYESSGNKEIDASYVDPLGPQIERPKTAAKKRIDEDD
                     FADEELGDDLLPE"
     primer_bind     complement(3862..3914)
                     /label=Rfrag-cib1-rfp-ift88a
     promoter        complement(3887..3904)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3909..4043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(4156..4174)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4195..4211)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4219..4235
                     /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)."
     promoter        complement(4243..4273)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4288..4309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4597..5185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5356..6216)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6217..6321)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6347..6802)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6880..7857
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg tcagatccgc tagcgccacc atgaatggag ctataggagg
      121 tgaccttttg ctcaattttc ctgacatgtc ggtcctagag cgccaaaggg ctcacctcaa
      181 gtacctcaat cccacctttg attctcctct cgccggcttc tttgccgatt cttcaatgat
      241 taccggcggc gagatggaca gctatctttc gactgccggt ttgaatcttc cgatgatgta
      301 cggtgagacg acggtggaag gtgattcaag actctcaatt tcgccggaaa cgacgcttgg
      361 gactggaaat ttcaaggcag cgaagtttga tacagagact aaggattgta atgaggcggc
      421 gaagaagatg acgatgaaca gagatgacct agtagaagaa ggagaagaag agaagtcgaa
      481 aataacagag caaaacaatg ggagcacaaa aagcatcaag aagatgaaac acaaagccaa
      541 gaaagaagag aacaatttct ctaatgattc atctaaagtg acgaaggaat tggagaaaac
      601 ggattatatt catgtaccgg tcgccacggc caccatggtg tctaagggcg aagagctgat
      661 taaggagaac atgcacatga agctgtacat ggagggcacc gtgaacaacc accacttcaa
      721 gtgcacatcc gagggcgaag gcaagcccta cgagggcacc cagaccatga gaatcaaggt
      781 ggtcgagggc ggccctctcc ccttcgcctt cgacatcctg gctaccagct tcatgtacgg
      841 cagcagaacc ttcatcaacc acacccaggg catccccgat ttctttaagc agtccttccc
      901 tgagggcttc acatgggaga gagtcaccac atacgaagac gggggcgtgc tgaccgctac
      961 ccaggacacc agcctccagg acggctgcct catctacaac gtcaagatca gaggggtgaa
     1021 cttcccatcc aacggccctg tgatgcagaa gaaaacactc ggctgggagg ccaacaccga
     1081 gatgctgtac cccgctgacg gcggcctgga aggcagaacc gacatggccc tgaagctcgt
     1141 gggcgggggc cacctgatct gcaacttcaa gaccacatac agatccaaga aacccgctaa
     1201 gaacctcaag atgcccggcg tctactatgt ggaccacaga ctggaaagaa tcaaggaggc
     1261 cgacaaagag acctacgtcg agcagcacga ggtggctgtg gccagatact gcgacctccc
     1321 tagcaaactg gggcacaaac ttaatggcat ggacgagctg tacaagactc gagctcaagc
     1381 ttcgaattca atgatggaaa atgttcatct ggcaccagaa acagatgagg acgaccttta
     1441 ctctggtttc aacgactaca acccagccta tgatactgag gagttggaga atgacacggg
     1501 ttttcagcaa gcagtgagaa ccagtcatgg cagaagacct cccgtaactg ctaaaatacc
     1561 aagcacagca gtcagcagac ccatagctac tggatatggg tccaagacat ccctgacatc
     1621 atcaatggga agaccgatga cagggacaat tcaggatgga gttgctcgac cgatgacagc
     1681 agtgagagca gctggctttt ccaaggcagc tttgcgaggc tctgcatttg acccccttgg
     1741 tcagtccagg ggccctgctc ctcctttgga agccaaaaat gaagacagtc cagaagaaaa
     1801 aattagacag ttggagaaga aagtaaatga gttggtggaa gaaagctgta ttgccaacag
     1861 ttgtggagac ttaaaactgg ccttagagaa ggcaaaagat gcaggaagaa aagagagagt
     1921 cctggtgaga caacgagagc aagtcacaag tccagaaaat atcaacttgg acttaaccta
     1981 ctccgttctt ttcaacttgg ccagtcagta ttctgctaat gaaatgtatg cagaagcact
     2041 gaacacatac caagttatag tgaaaaacaa gatgtttagc aatgcaggaa gactgaaagt
     2101 gaatatggga aatatttatt taaggcagag aaattattcc aaagccatta agttctaccg
     2161 aatggcctta gatcagatcc caagtgtcca taaagaaatg aggatcaaaa taatgcagaa
     2221 cattggaatc acatttataa aaactggtca gtactcagat gccatcaact catttgagca
     2281 catcatgagc atggccccga gcctgaaggc aggcttcaac ctcatcctta gttgttttgc
     2341 cattggagat cgggagaaaa tgaagaaggc attccaaaaa ttaattgctg ttccgttaga
     2401 aattgatgaa gatgataaat atatctcccc aagtgatgat ccccatacta acttgctgat
     2461 tgaagctata aaaaatgacc accttaggca aatggaacgt gaaaggaaag ccatggcaga
     2521 aaaatacatc atgacagctg caaaactcat tgctcctgtg atcgaagcgt cttttgctgt
     2581 gggttataac tggtgtgtgg aagtagtgaa agcctctcag tatgtggagc tggccaacga
     2641 cctggagatt aacaaagcaa ttacttactt gagacaaaag gactttaacc aagctgtaga
     2701 cacattgaaa atgtttgaga ggaaggacag tagagtgaag agtgcagctg cgaccaacct
     2761 ctcgttcctg tattatctgg aaaacgaatt tgcccaagcc agcagctatg cagatttagc
     2821 tgtgaactcg gatagatata acccgtcagc tctcactaat aaaggggaca cagtttttgc
     2881 aaatggtgat tatgagaagg cagctgagtt ctataaagag gccctgagaa acgactcttc
     2941 ctgtactgaa gcactgtata acattggcct cacctataag aagctgaacc gtctggatga
     3001 agccctggac tccttcctga aactgcacgc gatccttcgg aacagcgctc aagttctttg
     3061 ccagatagca aacatatatg agttaatgga agatcccaat caagctattg aatggctgat
     3121 gcagttgatc agcgttgttc caactgattc ccaagctctg tctaaactgg gagagttata
     3181 cgatagtgag ggagacaagt cccaggcatt ccaatattac tatgagtcat acaggtattt
     3241 cccttctaac attgaagtca ttgaatggct tggagcttat tacattgata cacagttctg
     3301 cgagaaagcc attcagtact ttgaaagagc ttcccttata caacccacac aagtgaagtg
     3361 gcagctgatg gtagctagct gctttagaag aagtggtaac taccaaaagg cattagatac
     3421 ttataaagag attcacagga aatttccggg gaatgttgaa tgtttgcgtt tcttggttcg
     3481 tctctgcaca gatattggat taaaagaagt tcaagaatat gccacaaaac tcaagaggtt
     3541 ggaaaaaatg aaagaaatga gggaacagcg cataaaatcg ggcagagaca gcagcggggg
     3601 ttctcgcagc aaacgagagg ggagtgctgg cagtgacagt ggccagaaca atagtgccag
     3661 tagtaagagt gaacgactga gtgccaagct cagagctttg cctgggacag atgaacctta
     3721 tgaaagtagc ggtaacaaag aaatagatgc ctcctacgtg gatccactgg gccctcagat
     3781 agaaagacca aaaactgcag ccaagaaaag aattgacgag gatgattttg ctgatgaaga
     3841 gttaggagat gacttgctcc cagaataagc ggcctgcagt ctagaactat agtgagtcgt
     3901 attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     3961 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4021 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4081 agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt
     4141 acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     4201 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag
     4261 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4321 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4381 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4441 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4501 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4561 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4621 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4681 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4741 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4801 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4861 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4921 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4981 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5041 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5101 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5161 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5221 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5281 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5341 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5401 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5461 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5521 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5581 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     5641 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     5701 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5761 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5821 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5881 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5941 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6001 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6061 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6121 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6181 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6241 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6301 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     6361 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     6421 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     6481 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     6541 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     6601 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     6661 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     6721 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6781 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     6841 cgatcggtgc gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata
     6901 tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata
     6961 tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact
     7021 tggcaccatg ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac
     7081 ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa
     7141 ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca
     7201 atatccaata tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc
     7261 cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa
     7321 tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc actcccccat
     7381 tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg
     7441 acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg
     7501 cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt
     7561 catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta
     7621 catcaatatc tattaatagt aacttggcaa gtacattact attggaagta cgccagggta
     7681 cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta
     7741 catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa
     7801 atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
