LOCUS       3xflag-Halo-Atg5        7118 bp DNA     circular SYN 04-OCT-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7118)
  AUTHORS   IQ
  TITLE     Direct Submission
  JOURNAL   Exported Aug 11, 2023 from SnapGene Viewer 7.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7118
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     mobile_element  403..627
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
                     /note="color: #ffe4c4"
     CDS             complement(694..1227)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /note="color: #ccffcc"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(1416..1444)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
                     /note="This reverse directional feature has 3 segments:
                        1: 1416 .. 1421 / #ffffff / -10
                        2: 1422 .. 1438 / #ffffff
                        3: 1439 .. 1444 / #ffffff / -35"
     polyA_signal    1953..2001
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
                     /note="color: #a6acb3"
     promoter        complement(2137..2246)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
                     /note="color: #ffffff; direction: LEFT"
     promoter        2270..2361
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(3047..3098)
                     /label=Vector.FOR
                     /note="color: black; sequence: 
                     CGTCATGGTCTTTGTAGTCCATtcttccaggagttaaagcagtacatacagg; 
                     added: 2019-10-04"
     primer_bind     3077..3176
                     /label=Fragment.FOR
                     /note="color: black; sequence: 
                     atggactacaaagaccatgacggtgattataaagatcatgatatcgattacaaggatga
                     cgatgacaagggttccatggcagaaatcggtactggctttc; added: 
                     2019-10-04"
     CDS             3077..3145
                     /codon_start=1
                     /label=3xFlag
                     /note="color: #cc99ff"
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     3077..3101
                     /label=oJS790
                     /note="color: black; sequence: atggactacaaagaccatgacggtg"
     primer_bind     3077..3099
                     /label=Fragment.FOR
                     /note="color: black; sequence: 
                     ggatcatcgataccatggactacaaagaccatgacgg"
     CDS             3152..4042
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /note="color: #cc99b2"
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     primer_bind     complement(3211..3229)
                     /label=ojs1086
                     /note="color: black; sequence: atcgacgtagtgcatgcgc"
     primer_bind     complement(4024..4062)
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     agatcctcagtggttggctcGCCGGAAATCTCGAGCGTC; added: 2019-10-04"
     primer_bind     4043..4066
                     /label=Vector.REV
                     /note="color: black; sequence: gagccaaccactgaggatctgtac; 
                     added: 2019-10-04"
     CDS             4055..4075
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /note="color: #cc99b2"
                     /translation="EDLYFQS"
     primer_bind     complement(4082..4099)
                     /label=Primer 2
                     /note="color: black; sequence: 
                     gatcagtttcGGTACCGGAAGCGATCGC"
     primer_bind     complement(4467..4490)
                     /label=ATG5_RHA_R2
                     /note="color: black; sequence: gaaggcaccatagctgatgattcc"
     polyA_signal    4500..4634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     mobile_element  4663..4828
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
                     /note="color: #ffe4c4"
     rep_origin      5012..5467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5494..5598
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5599..6459
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5599 .. 5667 / #ccffcc / signal sequence
                        2: 5668 .. 6459 / #ccffcc
                     Cleavage site after base 5668"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      join(6630..7118,1..100)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
       61 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      121 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      181 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
      241 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
      301 tctccttacg catctgtgcg gtatttcaca ccgcagacca gccgcgtaac ctggcaaaat
      361 cggttacggt tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag
      421 tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata
      481 gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg
      541 ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg
      601 gccaagggca tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc
      661 ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc
      721 aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc
      781 accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag
      841 gagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc
      901 atagatatag atctcactac gcggctgctc aaacctgggc agaacgtaag ccgcgagagc
      961 gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa
     1021 gttcccgagg taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc
     1081 acgaccgaaa agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca
     1141 tgtgcgaatg atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg
     1201 cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca
     1261 cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat
     1321 aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg
     1381 accagttgcg tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg
     1441 tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca
     1501 gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca
     1561 cgcatcgtca ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc
     1621 tgccctggct tcaggagatc ggtagacctc ggccgtcgcg gcgcttgccg gtggtgctga
     1681 ccccggatga agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc
     1741 aggactctag ctatagttct agtggttggc ctacgtaccc gtagtggcta tggcagggct
     1801 tgccgccccg acgttggctg cgagccctgg gccttcaccc gaacttgggg gttggggtgg
     1861 ggaaaaggaa gaaacgcggg cgtattggtc ccaatggggt ctcggtgggg tatcgacaga
     1921 gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac ccgtgcgttt
     1981 tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg
     2041 tttcagttag cctcccccat ctcccggtac cgcatgctat gcatcagctg ctagcaccat
     2101 ggctcgagat cccgggtgat caagtcttcg tcgagtgatt gtaaataaaa tgtaatttac
     2161 agtatagtat tttaattaat atacaaatga tttgataata attcttattt aactataata
     2221 tattgtgttg ggttgaatta aaggtccgta tactccggaa tattaataga tcatggagat
     2281 aattaaaatg ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt
     2341 gtaataaaaa aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga
     2401 tcccggtccg aagcgcgcgg aattcaaagg cctacgtcga cgagctcact agtcgcggcc
     2461 gctttcgaat ctagagcctg cagtctcgac aagcttgtcg agaagtacta gaggatcata
     2521 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     2581 ctgaacctga aacataaaat gaatgcaatt gttgttgttg gggcaggggt aaatgcgaga
     2641 atgatccttt ggagttggac tgtcaggatt cacagggtat aggtgaagct cctctttagg
     2701 taagcagtag actcttctgg gcttgaaaga ctgatgcaga tacagtttca ttttgactgt
     2761 ggttcagtac ttaaatatgc aatgaatatt actggcatat gtaattttga tctgattcct
     2821 ttacataagt aataatttta catttagaat ttgttgtact cttttctttg aatttatact
     2881 aaagattggt gatcatcgtt tttttttttt gaagtggata atctttatgg catggatttt
     2941 ttaaaataat cttttgtttt ctagcaggtt cattcatcgt tgcaaggatc tgactaatgc
     3001 tcttcatttt attttcagtt tggctttggt tgaaataaga atttagcctg tatgtactgc
     3061 tttaactcct ggaagaatgg actacaaaga ccatgacggt gattataaag atcatgatat
     3121 cgattacaag gatgacgatg acaagggttc catggcagaa atcggtactg gctttccatt
     3181 cgacccccat tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg
     3241 cgatggcacc cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa
     3301 catcatcccg catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg
     3361 caaatccgac aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc
     3421 cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc
     3481 tctgggtttc cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga
     3541 gttcatccgc cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca
     3601 ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga
     3661 gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg
     3721 cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc
     3781 aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca
     3841 ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc
     3901 cgaagccgct cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg
     3961 tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct
     4021 gtcgacgctc gagatttccg gcgagccaac cactgaggat ctgtactttc agagcgataa
     4081 cgcgatcgct tccggtacca cagatgacaa agatgtgctt cgagatgtgt ggtttggacg
     4141 aattccaact tgtttcacgc tatatcagga tgagataact gaaagggaag cagaaccata
     4201 ctatgtaagt acattgattg tggcaaatgg gatatttaac cttgtccatt tttgtaagaa
     4261 aaagctgtaa gaacttggag gccattatgt aacgttttag atacagctaa ttcatgctta
     4321 ttttggaaaa aaaattaaca cagataagca taaagaagga aataatcacc tctaatgcta
     4381 ccactcagag gacaaaacca ctgtcagcat gttgtcatga tgcgttctgg actttgtttt
     4441 ccttacatgt atgtgggtat gttaatggaa tcatcagcta tggtgccttc ttgactttca
     4501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4621 atcatgtctg gatctgatca ctgcttgagc ctaggagatc cgaaccagat aagtgaaatc
     4681 tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc tacacccagt
     4741 tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt ccacccctcc
     4801 cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt atgtttttaa
     4861 tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt ttctctgtca
     4921 cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga ctgaatatca
     4981 acgcttattt gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     5041 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5101 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5161 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5221 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5281 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5341 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5401 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5461 acaatttcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5521 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5581 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5641 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5701 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5761 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5821 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5881 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5941 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6001 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6061 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6121 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6181 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6241 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6301 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6361 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6421 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6481 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6541 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6601 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6661 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6721 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     6781 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6841 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6901 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6961 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7021 cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7081 agggtcggaa caggagagcg cacgagggag cttccagg
//
