LOCUS       pBP-mCh-Rab7-S72        8411 bp DNA     circular SYN 17-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8411)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 17, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    join(1..729,730..1353)
                     /locus_tag="mCh-Rab7"
                     /label=mCh-Rab7
     CDS             join(1..729,730..1353)
                     /codon_start=1
                     /label=mCh-Rab7
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKSGLRSRAMTSRKKVLLKVIILGDSGVGKTSLMNQYVNKKFSNQYK
                     ATIGADFLTKEVMVDDRLVTMQIWDTAGQERFQSLGVAFYRGADCCVLVFDVTAPNTFK
                     TLDSWRDEFLIQASPRDPENFPFVVLGNKIDLENRQVATKRAQAWCYSKNNIPYFETSA
                     KEAINVEQAFQTIARNALKQETEVELYNEFPEPIKLDKNDRAKASAESCSC"
     CDS             1..708
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     protein_bind    complement(1354..1378)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1521..1576
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     enhancer        1931..2310
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2311..2514
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2547..3146
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3190..3414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3893..3911)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3918..3934)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4076..4531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4557..4661
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4662..4730,4731..5522)
                     /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"
     rep_origin      5693..6281
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6569..6590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6605..6610,6611..6628,6629..6635)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6643..6659
                     /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     6667..6683
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6704..6722
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    7182..7202
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        7203..8381
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          7434..8372
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     protein_bind    8381..8405
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      join(8406..8411,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
       61 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      121 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      181 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
      241 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
      301 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
      361 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
      421 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
      481 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
      541 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
      601 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
      661 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagtc cggactcaga
      721 tctcgagcta tgacctctag gaagaaagtg ttgctgaagg ttatcatcct gggagattct
      781 ggagtcggga agacatcact catgaaccag tatgtgaata agaaattcag caatcagtac
      841 aaagccacaa taggagctga ctttctgacc aaggaggtga tggtggatga caggctagtc
      901 acaatgcaga tatgggacac agcaggacag gaacggttcc agtctctcgg tgtggccttc
      961 tacagaggtg cagactgctg cgttctggta tttgatgtga ctgcccccaa cacattcaaa
     1021 accctagata gctggagaga tgagtttctc atccaggcca gtccccgaga tcctgaaaac
     1081 ttcccatttg ttgtgttggg aaacaagatt gacctcgaaa acagacaagt ggccacaaag
     1141 cgggcacagg cctggtgcta cagcaaaaac aacattccct actttgagac cagtgccaag
     1201 gaggccatca acgtggagca ggcgttccag acgattgcac ggaatgcact taagcaggaa
     1261 acggaggtgg agctgtacaa cgaatttcct gaacctatca aactggacaa gaatgaccgg
     1321 gccaaggcct cggcagaaag ctgcagttgc tgaacccagc tttcttgtac aaagtggtga
     1381 tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg ccctggctca
     1441 caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct
     1501 tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa
     1561 ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa
     1621 tgagtatttg gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg
     1681 ttggctataa agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca
     1741 tagaaaagcc ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc
     1801 tttaacatcc ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg
     1861 actactccca gtcatagctg tccctcttct cttatggaga tccctcgacc tgcagcccaa
     1921 gcttcgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     1981 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2041 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2101 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2161 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2221 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2281 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2341 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2401 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2461 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2521 ctaactagag aacccactgc gccaccatga ccgagtacaa gcccacggtg cgcctcgcca
     2581 cccgcgacga cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg
     2641 ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac
     2701 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg
     2761 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg
     2821 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc
     2881 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc
     2941 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg
     3001 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg
     3061 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt
     3121 gcatgacccg caagcccggt gcctgactcg agtctagagg gcccgtttaa acccgctgat
     3181 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     3241 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     3301 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     3361 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggctcgag
     3421 ttaattaacg agagcataat attgatatgt gccaaagttg tttctgactg actaataagt
     3481 ataatttgtt tctattatgt ataggttaag ctaattactt attttataat acaacatgac
     3541 tgtttttaaa gtacaaaata agtttatttt tgtaaaagag agaatgttta aaagttttgt
     3601 tactttatag aagaaatttt gagtttttgt ttttttttaa taaataaata aacataaata
     3661 aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac ttaatatcta
     3721 ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt caattttacg
     3781 catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa ataatagttt
     3841 ctaatttttt tattattcag cctgctgtcg tgaataccga gctccaattc gccctatagt
     3901 gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     3961 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4021 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg
     4081 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4141 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4201 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4261 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     4321 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4381 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     4441 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4501 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
     4561 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4621 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4681 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4741 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4801 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4861 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4921 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4981 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5041 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5101 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5161 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5221 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5281 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5341 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5401 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5461 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5521 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5581 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5641 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5701 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5761 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5821 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5881 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5941 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6001 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6061 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6121 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6181 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6241 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6301 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6361 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6421 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6481 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6541 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6601 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6661 ttcacacagg aaacagctat gaccatgatt acgccaagct cgaaattaac cctcactaaa
     6721 gggaacaaaa gctggtacct cgcgcgactt ggtttgccat tctttagcgc gcgtcgcgtc
     6781 acacagcttg gccacaatgt ggtttttgtc aaacgaagat tctatgacgt gtttaaagtt
     6841 taggtcgagt aaagcgcaaa tcttttttaa ccctagaaag atagtctgcg taaaattgac
     6901 gcatgcattc ttgaaatatt gctctctctt tctaaatagc gcgaatccgt cgctgtgcat
     6961 ttaggacatc tcagtcgccg cttggagctc ccgtgaggcg tgcttgtcaa tgcggtaagt
     7021 gtcactgatt ttgaactata acgaccgcgt gagtcaaaat gacgcatgat tatcttttac
     7081 gtgactttta agatttaact catacgataa ttatattgtt atttcatgtt ctacttacgt
     7141 gataacttat tatatatata ttttcttgtt atagatatca tcaactttgt atagaaaagt
     7201 tgggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
     7261 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
     7321 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
     7381 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
     7441 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
     7501 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
     7561 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
     7621 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
     7681 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
     7741 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
     7801 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
     7861 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
     7921 ctggtctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     7981 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     8041 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     8101 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     8161 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     8221 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     8281 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     8341 cagtggttca aagttttttt cttccatttc aggtgtcgtg acaagtttgt acaaaaaagc
     8401 aggctgccac c
//
