LOCUS       pNP-792_pCDNA3.1        7906 bp    DNA     circular SYN 13-MAY-2022
DEFINITION  New Assembly 7906bp ds-DNA circular SYN 13-MAY-2022.
ACCESSION
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2022 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7906
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    72..296
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      342..770
     promoter        784..1113
                     /note="SV40 enhancer and early promoter"
                     /standard_name="SV40 promoter"
     rep_origin      964..1099
     CDS             1180..1974
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="confers resistance to neomycin, kanamycin, and G418
                      (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
                     /standard_name="NeoR/KanR"
     polyA_signal    2148..2269
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2318..2334)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /standard_name="M13 rev"
     protein_bind    2342..2358
                     /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)."
                     /standard_name="lac operator"
     promoter        complement(2366..2396)
                     /note="promoter for the E. coli lac operon"
                     /standard_name="lac promoter"
     protein_bind    2411..2432
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                      of cAMP."
                     /standard_name="CAP binding site"
     rep_origin      complement(2720..3305)
     CDS             complement(3476..4336)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /standard_name="AmpR"
     promoter        complement(4337..4441)
                     /gene="bla"
                     /standard_name="AmpR promoter"
     enhancer        4707..5086
                     /note="human cytomegalovirus immediate early enhancer"
                     /standard_name="CMV enhancer"
     promoter        5087..5290
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /standard_name="CMV promoter"
     promoter        5335..5353
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /standard_name="T7 promoter"
     primer_bind     5364..5401
                     /standard_name="NAPH-p152F"
     misc_feature    5384..5389
                     /standard_name="Kozak"
     misc_feature    5387..5443
                     /standard_name="ADGRL3 SP"
     CDS             5444..5989
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine
                     derivatives"
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAV
                     EVPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKL
                     LKVVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEG
                     GLAVKEWLLAHEGHRLGKPGLG"
                     /standard_name="SNAP-tag(R)"
     misc_feature    complement(5972..5999)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="NAPH-p396R"
     primer_bind     5980..6008
                     /standard_name="NAPH-p397F"
     misc_feature    5990..6016
                     /standard_name="linker"
     CDS             6017..6040
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase
                      cleavage site"
                     /translation="DYKDDDDK"
                     /standard_name="FLAG"
     misc_feature    6041..7903
                     /standard_name="ADGRL3-CTF"
     misc_feature    complement(7877..20)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="NAPH-p398R"
     misc_feature    complement(7886..20)
                     /note="Geneious type: primer_bind_reverse"
                     /standard_name="NAPH-p203R"
ORIGIN      
        1 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg
       61 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
      121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
      181 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
      241 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
      301 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
      361 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
      421 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
      481 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
      541 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
      601 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
      661 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
      721 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
      781 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
      841 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
      901 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
      961 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     1021 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     1081 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     1141 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     1201 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     1261 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     1321 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     1381 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     1441 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     1501 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     1561 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     1621 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     1681 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     1741 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     1801 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     1861 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     1921 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     1981 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     2041 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     2101 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
     2161 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     2221 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
     2281 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     2341 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     2401 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     2461 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     2521 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2581 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     2641 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     2701 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     2761 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     2821 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     2881 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2941 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     3001 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     3061 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     3121 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     3181 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     3241 ccaccgctgg tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3301 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3361 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3421 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3481 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3541 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3601 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3661 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3721 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3781 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3841 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3901 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3961 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     4021 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     4081 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     4141 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     4201 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4261 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4321 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4381 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4441 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     4501 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     4561 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     4621 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     4681 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     4741 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     4801 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     4861 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     4921 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     4981 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     5041 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     5101 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     5161 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     5221 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     5281 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     5341 gactcactat agggagaccc aagctggcta gcgtttaaac ttagccatgt ggccaccaca
     5401 gctactcatc ctcacgatgc ttttagcacc tgtagttcat ggtatggaca aagactgcga
     5461 aatgaagcgc accaccctgg atagccctct gggcaagctg gaactgtctg ggtgcgaaca
     5521 gggcctgcac cgtatcatct tcctgggcaa aggaacatct gccgccgacg ccgtggaagt
     5581 gcctgcccca gccgccgtgc tgggcggacc agagccactg atgcaggcca ccgcctggct
     5641 caacgcctac tttcaccagc ctgaggccat cgaggagttc cctgtgccag ccctgcacca
     5701 cccagtgttc cagcaggaga gctttacccg ccaggtgctg tggaaactgc tgaaagtggt
     5761 gaagttcgga gaggtcatca gctacagcca cctggccgcc ctggccggca atcccgccgc
     5821 caccgccgcc gtgaaaaccg ccctgagcgg aaatcccgtg cccattctga tcccctgcca
     5881 ccgggtggtg cagggcgacc tggacgtggg gggctacgag ggcgggctcg ccgtgaaaga
     5941 gtggctgctg gcccacgagg gccacagact gggcaagcct gggctgggtg gcggctctgg
     6001 cggcagcggc ggaagcgatt ataaggacga cgacgacaag accaactttg cagtattgat
     6061 ggcacatgtg gaagttaagc acagcgatgc agtccacgat cttcttctgg atgtgatcac
     6121 gtgggttggc atcttgctgt cccttgtctg tctcctgatt tgcatcttca cattctgctt
     6181 cttccgggga ctccagagtg accgcaacac cattcacaag aacctctgca tcagcctgtt
     6241 cgtggcagaa ctgctcttcc tgattggaat caacagaacc gaccaaccaa ttgcctgtgc
     6301 agtgttcgcg gctcttttgc atttcttctt cttggcggcc ttcacctgga tgtttctaga
     6361 aggggtgcag ctgtatatca tgctggtgga ggtctttgag agcgagcatt cccgtaggaa
     6421 gtacttttat ctggtcggct atgggatgcc cgcgctcatc gtggccgttt ctgcggccgt
     6481 cgactacagg agctacggaa cagacaaagt atgttggctt cgccttgaca cctacttcat
     6541 ttggagtttt ataggaccgg cgaccttgat aattatgctg aatgtcatct tccttgggat
     6601 tgctttatac aaaatgtttc accatactgc catactgaaa cctgaatcag gctgtcttga
     6661 caatatcaac tatgaggata acagaccctt catcaagtca tgggttatag gtgcaatagc
     6721 tctgctctgc ctgttaggat tgacctgggc ctttggactc atgtatatta atgaaagcac
     6781 agtcatcatg gcgtatctct tcaccatttt caattctcta cagggaatgt ttatattcat
     6841 tttccactgt gttctacaga agaaggtacg gaaagagtat gggaaatgcc tccgcacgca
     6901 ttgctgtagt gggaaaagca cggagagttc gattggctca gggaaaacat ctggttctcg
     6961 aactccagga cggtactcca caggctcgca gagccggatt cggagaatgt ggaatgacac
     7021 tgtccgaaag cagtcagagt cgtccttcat caccggagac ataaacagct cagcatcgct
     7081 caacagagag ccctacagag agacaaaggg gcttctgaac aatgccaggg atacaagtgt
     7141 catggatact ctaccactga atggtaacca tggcaacagt tacagcattg ctggcggcga
     7201 atacctgagc aactgtgtgc aaattataga ccgtggctat aaccacaacg agaccgccct
     7261 agagaaaaag atcctaaagg aactcacttc caactatatc ccttcatacc tgaacaacca
     7321 cgagcgctcc agcgaacaga accggaatat gatgaacaaa ctggtgaaca acttaggcag
     7381 cgggagtgaa gatgacgcca tcgtcctgga tgacgccgcg tcctttaacc acgaggagag
     7441 tctgggcctg gaactcattc acgaggaatc cgatgctccc ttgctgcccc cgagggttta
     7501 ctccacggat aaccaccagc cacaccatta cagcaggagg cggttccccc aggaccacag
     7561 cgagagcttc ttccctctgc taaccgacga gcacacagaa gacctgcagt caccgcacag
     7621 ggactctctg tacaccagca tgccagccct ggccggcgtg cccgctgcag acagtgtgac
     7681 caccagcacc cagaccgaag ccgcagcggc caagggtggt gacgccgaag atgtttacta
     7741 caaaagcatg ccaaacctgg gctccagaaa ccacgtgcac ccgctgcacg cctactacca
     7801 gctggggcgc ggcagcagcg acggattcat agttcctcct aacaaagatg gggcctctcc
     7861 ggaggggact tccaaaggac ccgcgcactt ggtcactagt ctataa
//
