LOCUS       pAc5.1-V5-HisB-O        7912 bp DNA     circular SYN 09-SEP-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7912)
  AUTHORS   Matthew A Birk
  TITLE     Direct Submission
  JOURNAL   Exported Jun 8, 2023 from SnapGene 6.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7912
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     primer_bind     2543..2561
                     /label=MB0208
     primer_bind     2584..2630
                     /label=MB0205
     misc_feature    2584..2603
                     /label=EcoRI Gibson overlap
     misc_feature    2606..2612
                     /label=Drosophila Kozak sequence
     CDS             2613..4283
                     /codon_start=1
                     /label=Obimac KHC (1-558)
                     /translation="DASSECNIKVICRVRPLNDAEERAGSKFILKFPTDDSISIAGKVF
                     VFDKVLKPNVSQEYVYNVGAQPIVADVLSGYNGTIFAYGQTSSGKTHTMEGVLDKPSMH
                     GIIPRIVQDIFNYIYGMDENLEFHIKISYYEIYLDKIRDLLDVTKTNLSVHEDKNRVPY
                     VKGATERFVSSPEEVMEVIDEGKNNRHVAVTNMNEHSSRSHSVFLINVKQENVETQKKL
                     SGKLYLVDLAGSEKVSKTGAEGAVLDEAKNINKSLSALGNVISALADGNKSHVPYRDSK
                     LTRILQESLGGNARTTMVICCSPASYNESETKSTLLFGQRAKTIKNVVSVNEELTADEW
                     KRRYEKEKEKCTKLKATLARLESELQKWRTGQPVSVDDQVDLKADIPAESPATSTTSLA
                     GNMVVSMNEGDRSQLEEERLKLYQQLDDKDDEINNQSQLIEKLNEQMMEQEELISQSRK
                     DYENLQQEMSRIQSDNESAKDEVKEVLQALEELAMNYDQKSQEVENKNKENENLSDELS
                     QKLSTLNSLQNELDQLKDSSMHHRKRVTDMMINLLKDLGD"
     primer_bind     3307..3325
                     /label=MB0150
     mat_peptide     3453..3455
                     /label=K282R
     primer_bind     4037..4055
                     /label=MB0151
     misc_feature    4279..4298
                     /label=GA overlap for eGFP or HaloTag
     primer_bind     4279..4297
                     /label=MB0152
     CDS             4299..5189
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(5173..5212)
                     /label=MB0206
     primer_bind     complement(5173..5190)
                     /label=MB0203
     misc_feature    5195..5212
                     /label=BstBI Gibson overlap
     CDS             5197..5238
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             5248..5265
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5338..5472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5371..5391)
                     /label=MB0209
     rep_origin      complement(6094..6682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6853..7713)
                     /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"
     promoter        complement(7714..7818)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 tacctactag tccagtgtgg tggatcaaca tggatgcatc ttcagaatgc aatattaaag
     2641 tcatatgccg ggtgagacct ctcaatgatg cagaagagcg agcaggctcc aagtttatac
     2701 tcaaatttcc taccgatgat tctataagta tagctggtaa agtgttcgtt tttgataaag
     2761 tgctcaagcc aaacgtaagt caagagtacg tctacaatgt tggcgcacag ccgattgtgg
     2821 ctgatgtcct tagtggatat aacgggacta ttttcgccta cggtcaaacc agttccggaa
     2881 agacgcacac aatggaagga gtccttgata aaccgtctat gcacggcatc attccgcgca
     2941 tagttcagga catattcaat tatatatacg ggatggatga gaacctagag ttccacataa
     3001 agatttccta ctatgaaata tacttggata aaataagaga tctcctcgat gtaactaaaa
     3061 cgaacttgtc agtgcacgag gacaaaaatc gcgtaccgta tgtcaaggga gccactgaga
     3121 gatttgtcag tagcccggag gaggttatgg aagttattga tgagggtaaa aataaccggc
     3181 atgtcgctgt aaccaacatg aacgagcaca gttcaaggtc tcactccgtg tttttgatca
     3241 acgtaaaaca ggagaacgtc gaaacgcaaa agaaactttc cggaaaactg taccttgtgg
     3301 accttgccgg atcagaaaag gtgtctaaaa cgggcgcgga aggagcggtt ctcgacgagg
     3361 caaaaaacat taataagagt cttagcgcac tggggaatgt tatcagcgcg ctcgctgatg
     3421 ggaacaagag ccatgtcccc taccgagata gtaaactgac gcggatcttg caagaatctc
     3481 ttggaggcaa tgcaaggaca accatggtca tatgctgtag tcctgcttcc tacaatgaaa
     3541 gcgaaaccaa atcaacactc ctgttcggtc aaagggcgaa aactatcaag aatgttgtca
     3601 gtgtaaacga agaattgacg gccgacgagt ggaagcgcag atatgagaag gaaaaggaga
     3661 aatgcacgaa attgaaagct acgctcgcca gacttgaatc tgaactgcaa aaatggcgaa
     3721 caggccaacc cgtatctgtt gatgaccagg tggatctcaa ggctgatata cctgcggaat
     3781 cccccgcgac aagtaccaca tccttggctg ggaatatggt ggtatcaatg aatgaaggag
     3841 accgctcaca actggaggag gagcgattga agctgtatca gcaactcgac gacaaggatg
     3901 acgagatcaa taatcagagc cagctcatag agaaattgaa tgaacaaatg atggagcaag
     3961 aagaactcat atcccaatca cggaaggatt acgagaatct tcagcaagag atgagccgaa
     4021 tccagagcga caatgagtct gctaaggacg aagtgaaaga agtgctgcaa gcattggagg
     4081 aattggcgat gaactatgac caaaagtccc aggaagtgga aaacaagaat aaggagaatg
     4141 agaatctctc cgatgagttg tcccaaaaac ttagtaccct taattcactc cagaatgaac
     4201 tggatcaact aaaggacagc agtatgcatc accgcaagcg cgtgaccgac atgatgatca
     4261 accttctcaa ggacctggga gatggaggtg gaggttcaat ggcggaaata ggcacgggct
     4321 ttccgtttga ccctcactac gttgaggtac ttggggaacg gatgcattac gtagatgtag
     4381 ggccaagaga cgggacgccg gtactcttcc tgcacgggaa tccaacgagc agctacgtgt
     4441 ggagaaatat catcccgcat gtcgccccga ctcataggtg tatagctcca gatttgattg
     4501 gcatgggaaa atcagataaa cctgatcttg gttacttttt tgacgaccat gtgaggttta
     4561 tggacgcatt catagaggcg cttggtctcg aggaggttgt acttgtcata catgactggg
     4621 gttctgcact cggcttccac tgggcaaagc gcaatcctga aagggtaaaa ggcatcgcat
     4681 ttatggagtt tattcggcct attcccacgt gggatgaatg gccggaattt gctagagaga
     4741 cgtttcaggc atttcgcacc acggatgtag ggcggaaact tattattgac caaaatgtat
     4801 tcatagaggg gactcttccg atgggtgttg tgaggccgct tactgaggtg gaaatggatc
     4861 actatagaga gcctttcctt aatccagtcg accgagagcc tctgtggcgg ttccccaatg
     4921 aacttcccat agcgggagaa ccggccaaca ttgtcgcgct ggtcgaggaa tacatggact
     4981 ggctgcatca gagccccgta ccaaaattgt tgttttgggg aacgccaggg gtgctgatac
     5041 ctcccgcaga ggcggccaga ctcgcgaaat ccctccccaa ttgtaaagcc gttgatatag
     5101 ggcccggttt gaatcttttg caagaggata atccagattt gataggttct gagattgcga
     5161 gatggttgag cacactggaa atctcaggct aacgaaggta agcctatccc taaccctctc
     5221 ctcggtctcg attctacgcg taccggtcat catcaccatc accattgagt ttaaacccgc
     5281 tgatcagcct cgactgtgcc ttctaaggcc tgagctcgct gatcagcctc gatcgaggat
     5341 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     5401 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     5461 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     5521 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtatggctga ttatgatcag
     5581 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     5641 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     5701 ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg
     5761 cctgtcgctt gcggtattcg gaatcttgca cgccctcgct caagccttcg tcactggtcc
     5821 cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct
     5881 gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg gccttcccca ttatgattct
     5941 tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga
     6001 tgacgaccat cagggacagc ttcaaggatc gctcgcggct cttaccagcc agcaaaaggc
     6061 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     6121 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     6181 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6241 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcaat gctcacgctg
     6301 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6361 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6421 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6481 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     6541 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6601 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6661 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6721 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6781 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6841 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6901 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6961 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     7021 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7081 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7141 tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg
     7201 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7261 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7321 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7381 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7441 gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac
     7501 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7561 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7621 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7681 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7741 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7801 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7861 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gt
//
