LOCUS       62056_2365        6978 bp DNA     circular SYN 27-JUN-2020
DEFINITION  Cloning vector pAAV-EF1A-mRuby2-Gephyrin.FingR-IL2RGTC, complete 
            sequence.
ACCESSION   MT612428
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6978)
  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  JOURNAL   Unpublished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 6978)
  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 6978)
  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            "Unpublished"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..6978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    228..249
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        264..294
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    302..318
                     /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)."
     primer_bind     326..342
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    379..564
                     /label=5' ITR
                     /note="5' ITR"
     misc_feature    575..586
                     /label=IL2RG binding site
                     /note="IL2RG binding site"
     promoter        598..1776
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1793..1811
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1844..2554
                     /label=mRuby2
                     /note="monomeric red fluorescent protein derived from
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
     CDS             2861..2884
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    2909..3433
                     /label=IL2RGTC
                     /note="IL2RGTC"
     misc_feature    3487..4075
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(4078..4094)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(4124..4245)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4348..4537
                     /label=3' ITR
                     /note="3' ITR"
     primer_bind     complement(4555..4571)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4713..5168
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5194..5298
                     /label=AmpR promoter
     CDS             5299..6156
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6330..6918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
       61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      301 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      361 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      421 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      481 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      541 atctacgtag ccatgctcta ggaagatcac gcgtcttcca cagagtgaat tccgtgaggc
      601 tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga
      661 ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat
      721 gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta
      781 gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg
      841 tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact
      901 tccacctggc tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag
      961 agttcgaggc cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct
     1021 gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc
     1081 gataagtctc tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa
     1141 gatagtcttg taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg
     1201 ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg
     1261 cggccaccga gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc
     1321 ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt
     1381 gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg
     1441 cggcgctcgg gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc
     1501 tcagccgtcg cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag
     1561 ttctcgagct tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag
     1621 tttccccaca ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc
     1681 tccttggaat ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg
     1741 gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa ttagcttggt actaatacga
     1801 ctcactatag ggagacccaa gctggctagg taagcttgcc accatggtgt ctaagggcga
     1861 agagctgatc aaggaaaata tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca
     1921 ccaattcaaa tgcacaggtg aaggagaagg caatccgtac atgggaactc aaaccatgag
     1981 gatcaaagtc atcgagggag gacccctgcc atttgccttt gacattcttg ccacgtcgtt
     2041 catgtatggc agccgtactt ttatcaagta cccgaaaggc attcctgatt tctttaaaca
     2101 gtcctttcct gagggtttta cttgggaaag agttacgaga tacgaagatg gtggagtcgt
     2161 caccgtcatg caggacacca gccttgagga tggctgtctc gtttaccacg tccaagtcag
     2221 aggggtaaac tttccctcca atggtcccgt gatgcagaag aagaccaagg gttgggagcc
     2281 taatacagag atgatgtatc cagcagatgg tggtctgagg ggatacactc atatggcact
     2341 gaaagttgat ggtggtggcc atctgtcttg ctctttcgta acaacttaca ggtcaaaaaa
     2401 gaccgtcggg aacatcaaga tgcccggtat ccatgccgtt gatcaccgcc tggaaaggtt
     2461 agaggaaagt gacaatgaaa tgttcgtagt acaacgcgaa cacgcagttg ccaagttcgc
     2521 cgggcttggt ggtgggatgg acgagctgta caagtctagg ggagctggcg ctggagcggg
     2581 tgcaggggct ggcctcgaag tcaaggaagc atcaccaacc agcatccaga tcagctgggg
     2641 caagtacaag gtcatggttc gctactaccg catcacctac ggtgaaactg gtggcaatag
     2701 ccctgtccag gaattcaccg tgcctggcag caagtccact gctaccatca gcagcctgaa
     2761 acctggtgtc gactatacca tcacggtgta cgccgtcacg atcgaccact ggaactacca
     2821 ggacccgatc ccgatctcca tcaactaccg caccggatcc gattacaagg atgacgacga
     2881 taagggtagc ggctccagta gatctgggtt tcagtgccag atttgcatgc gcaactttag
     2941 ccgcaaaagc accctgaccg atcatattcg cacccatacc ggcgaaaaac cgtttgcgtg
     3001 cgatatttgc ggccgcaaat ttgcggcgcg cagcacccgc accacccata ccaaaattca
     3061 taccggcagc cagaaaccgt ttcagtgccg catttgcatg cgcaacttta gccgcagcga
     3121 tagcctgagc aaacatattc gcacccatac cggcgaaaaa ccgtttgcgt gcgatatttg
     3181 cggccgcaaa tttgcgcagc gcagcaacct gaaagtgcat accaaaattc atctgcgcgg
     3241 cagccagctg atcgatggtg tcgacgctcc tgaacaacgt gaaggtgctt ctcaagtttc
     3301 tgtttctgtt acttttgaag atgttgctgt tctttttact cgtgatgaat ggaaaaaact
     3361 tgatctttct caacgttctc tttatcgtga agttatgctt gaaaattatt ctaatcttgc
     3421 ttctatggct taaaagcttg cggccgcgac tctagagtcg acctgcaggc atgcaagctt
     3481 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3541 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3601 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3661 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3721 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3781 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3841 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     3901 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3961 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4021 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     4081 taccgtcgac ctcgacccgg gcggccgctt cgagcagaca tgataagata cattgatgag
     4141 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     4201 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     4261 attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag caagtaaaac
     4321 ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg tagataagta
     4381 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4441 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4501 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     4561 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4621 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4681 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4741 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4801 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4861 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4921 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4981 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5041 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5101 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5161 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5221 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5281 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5341 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5401 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5461 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5521 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5581 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5641 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5701 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5761 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5821 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5881 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5941 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6001 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6061 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6121 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6181 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6241 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6781 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6841 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6901 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6961 tggccttttg ctcacatg
//
