LOCUS       Exported                7459 bp ds-DNA     circular SYN 19-APR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7459)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Apr 19, 2018 from SnapGene 4.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7459
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        93..472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        473..676
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        721..739
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    762..786
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      788..797
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             790..2040
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     protein_bind    complement(2042..2066)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2082..2102)
                     /label=PLAMP1-pmturquoise-BB.REV
     primer_bind     2085..2106
                     /label=plamp1-PMTURQUOISE-INS.FOR
     CDS             2086..2802
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2086..2089
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(2776..2797)
                     /label=plamp1-PMTURQUOISE-INS.REV
     primer_bind     2782..2807
                     /label=PLAMP1-pmturquoise-BB.FOR
     polyA_signal    2914..3138
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3184..3612
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3626..3955
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3806..3941
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4022..4816
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4990..5111
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5160..5176)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5184..5200
                     /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(5208..5238)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5253..5274
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5562..6150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6321..7181)
                     /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(7182..7286)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
       61 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      661 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
      721 taatacgact cactataggg agacccaagc tggctagatc aacaagtttg tacaaaaaag
      781 caggctacca tggcggcccc cggcagcgcc cggcgacccc tgctgctgct actgctgttg
      841 ctgctgctcg gcctcatgca ttgtgcgtca gcagcaatgt ttatggtgaa aaatggcaac
      901 gggaccgcgt gcataatggc caacttctct gctgccttct cagtgaacta cgacaccaag
      961 agtggcccta agaacatgac ctttgacctg ccatcagatg ccacagtggt gctcaaccgc
     1021 agctcctgtg gaaaagagaa cacttctgac cccagtctcg tgattgcttt tggaagagga
     1081 catacactca ctctcaattt cacgagaaat gcaacacgtt acagcgtcca gctcatgagt
     1141 tttgtttata acttgtcaga cacacacctt ttccccaatg cgagctccaa agaaatcaag
     1201 actgtggaat ctataactga catcagggca gatatagata aaaaatacag atgtgttagt
     1261 ggcacccagg tccacatgaa caacgtgacc gtaacgctcc atgatgccac catccaggcg
     1321 tacctttcca acagcagctt cagcagggga gagacacgct gtgaacaaga caggccttcc
     1381 ccaaccacag cgccccctgc gccacccagc ccctcgccct cacccgtgcc caagagcccc
     1441 tctgtggaca agtacaacgt gagcggcacc aacgggacct gcctgctggc cagcatgggg
     1501 ctgcagctga acctcaccta tgagaggaag gacaacacga cggtgacaag gcttctcaac
     1561 atcaacccca acaagacctc ggccagcggg agctgcggcg cccacctggt gactctggag
     1621 ctgcacagcg agggcaccac cgtcctgctc ttccagttcg ggatgaatgc aagttctagc
     1681 cggtttttcc tacaaggaat ccagttgaat acaattcttc ctgacgccag agaccctgcc
     1741 tttaaagctg ccaacggctc cctgcgagcg ctgcaggcca cagtcggcaa ttcctacaag
     1801 tgcaacgcgg aggagcacgt ccgtgtcacg aaggcgtttt cagtcaatat attcaaagtg
     1861 tgggtccagg ctttcaaggt ggaaggtggc cagtttggct ctgtggagga gtgtctgctg
     1921 gacgagaaca gcatgctgat ccccatcgct gtgggtggtg ccctggcggg gctggtcctc
     1981 atcgtcctca tcgcctacct cgtcggcagg aagaggagtc acgcaggcta ccagactatc
     2041 cacccagctt tcttgtacaa agtggttgat ctagcgttta ccggtatggt gagcaagggc
     2101 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     2161 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     2221 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     2281 tcctggggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     2341 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     2401 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     2461 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     2521 tactttagcg acaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     2581 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag
     2641 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     2701 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2761 accgccgccg ggatcactct cggcatggac gagctgtaca agggtaccga gctcggatcc
     2821 actagtccag tgtggtggaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta
     2881 gagggcccgt tgaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     2941 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     3001 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     3061 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     3121 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     3181 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     3241 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3301 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3361 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     3421 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3481 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     3541 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     3601 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3661 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3721 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3781 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3841 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     3901 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3961 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     4021 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4081 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     4141 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     4201 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     4261 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     4321 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     4381 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     4441 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     4501 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     4561 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4621 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4681 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4741 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4801 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4861 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4921 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4981 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5041 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5101 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5161 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5221 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5281 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5341 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5401 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6001 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6241 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6301 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6421 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6481 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6601 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7201 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7261 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     7321 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     7381 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     7441 gcaaaattta agctacaac
//
