LOCUS       62056_17335        7231 bp DNA     circular SYN 27-JUN-2020
DEFINITION  Cloning vector pMSCV-syn-PSD95.FingR-GFP, complete sequence.
ACCESSION   MT612433
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7231)
  AUTHORS   Bensussen S, Shankar S, Ching KH, Zemel D, Ta TL, Mount RA, Shroff 
            SN, Gritton HJ, Fabris P, Vanbenschoten H, Beck C, Man H-Y., Han X.
  TITLE     A viral toolbox of genetically encoded fluorescent synaptic tags
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7231)
  AUTHORS   Bensussen S, Shankar S, Ching KH, Zemel D, Ta TL, Mount RA, Shroff 
            SN, Gritton HJ, Fabris P, Vanbenschoten H, Beck C, Man H-Y., Han X.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JUN-2020) Biomedical Engineering, Boston University, 
            44 Cummington Mall, Boston, MA 02215, USA
REFERENCE   3  (bases 1 to 7231)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-JUN-2020) Biomedical Engineering, Boston University, 44 
            Cummington Mall, Boston, MA 02215, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7231
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             342..517
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             989..1405
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     promoter        1445..1892
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2202..2225
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             2250..2960
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELY"
     misc_feature    2976..3553
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             3592..4107
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     primer_bind     complement(4276..4292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4300..4316)
                     /label=lac operator
                     /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(4324..4354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4369..4390)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4678..5266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5440..6297)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6298..6402)
                     /label=AmpR promoter
ORIGIN
        1 ctgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc tagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatcttct agactgcaga gggccctgcg
     1441 tatgagtgca agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga
     1501 ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat
     1561 cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac
     1621 cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag
     1681 gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg
     1741 tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg
     1801 ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca
     1861 gcggaggagt cgtgtcgtgc ctgagagcgc agtcgagaac gcgtggtacc gagctcggat
     1921 ccgccgccac catgctcgaa gtcaaggaag catcaccaac cagcatccag atcagctggg
     1981 tgctccactt gcgccacgtt cgctactacc gcatcaccta cggtgaaact ggtggcaata
     2041 gccctgtcca ggaattcacc gtgcctggca gcaagtccac tgctaccatc agcggcctga
     2101 aacctggtgt cgactatacc atcacggtgt acgccgtcac gatcttcagc gcctaccgct
     2161 ccgcctggcc gccgatctcc atcaactacc gcaccggaac cgattacaag gatgacgacg
     2221 ataagggtag cggctccagt agatctgggg tgagcaaggg cgaggagctg ttcaccgggg
     2281 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2341 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2401 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2461 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2521 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2581 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2641 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2701 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2761 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2821 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2881 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2941 tcggcatgga cgagctgtac taaaagctta tcgataatca acctctggat tacaaaattt
     3001 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3061 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3121 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3181 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3241 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3301 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3361 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3421 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3481 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3541 tctccctttg ggcaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac
     3601 cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata
     3661 cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg
     3721 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     3781 tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     3841 ccccaaggac ctgaaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
     3901 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
     3961 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc
     4021 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
     4081 gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat
     4141 tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc
     4201 acatactcca atactcctga aatagttcat tatggacagc gcagaagagc tggggagaat
     4261 taattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4321 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4381 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4441 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     4501 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4561 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4621 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4681 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4741 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4801 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4861 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4921 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4981 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     5041 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     5101 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     5161 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     5221 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     5281 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     5341 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     5401 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     5461 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     5521 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5581 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5641 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5701 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5761 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5821 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5881 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5941 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     6001 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     6061 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     6121 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     6181 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     6241 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     6301 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6361 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     6421 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     6481 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     6541 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     6601 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     6661 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     6721 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     6781 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6841 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
     6901 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     6961 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     7021 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     7081 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
     7141 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
     7201 attttattta gtctccagaa aaagggggga a
//
