LOCUS       Exported                6975 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA-SADB19G
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6975)
  AUTHORS   Osakada F, Mori T, Cetin AH, Marshel JH, Virgen B, Callaway EM
  TITLE     New rabies virus variants for monitoring and manipulating activity 
            and gene expression in defined neural circuits.
  JOURNAL   Neuron. 2011 Aug 25;71(4):617-31.
  PUBMED    21867879
REFERENCE   2  (bases 1 to 6975)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6975
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      679..688
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(2294..2312)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2295..2312)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2332..2349)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2338..2562
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2608..3036
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2695..2714)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2905..2926
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3045..3065)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3050..3379
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3230..3365
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3292..3311
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3446..4240
                     /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"
     primer_bind     complement(3500..3519)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4110..4129
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4414..4535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4451..4470)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4505..4524
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4584..4600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4584..4600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4597..4619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4608..4624
                     /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(4632..4662)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4677..4698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4815..4832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4986..5574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5066..5085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5745..6605)
                     /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"
     primer_bind     6368..6387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6606..6710)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6785..6804)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccgc caccatggtt cctcaggctc tcctgtttgt accccttctg
      721 gtttttccat tgtgttttgg gaaattccct atttacacga taccagacaa gcttggtccc
      781 tggagtccga ttgacataca tcacctcagc tgcccaaaca atttggtagt ggaggacgaa
      841 ggatgcacca acctgtcagg gttctcctac atggaactta aagttggata catcttagcc
      901 ataaaagtga acgggttcac ttgcacaggc gttgtgacgg aggctgaaac ctacactaac
      961 ttcgttggtt atgtcacaac cacgttcaaa agaaagcatt tccgcccaac accagatgca
     1021 tgtagagccg cgtacaactg gaagatggcc ggtgacccca gatatgaaga gtctctacac
     1081 aatccgtacc ctgactaccg ctggcttcga actgtaaaaa ccaccaagga gtctctcgtt
     1141 atcatatctc caagtgtggc agatttggac ccatatgaca gatcccttca ctcgagggtc
     1201 ttccctagcg ggaagtgctc aggagtagcg gtgtcttcta cctactgctc cactaaccac
     1261 gattacacca tttggatgcc cgagaatccg agactaggga tgtcttgtga catttttacc
     1321 aatagtagag ggaagagagc atccaaaggg agtgagactt gcggctttgt agatgaaaga
     1381 ggcctatata agtctttaaa aggagcatgc aaactcaagt tatgtggagt tctaggactt
     1441 agacttatgg atggaacatg ggtctcgatg caaacatcaa atgaaaccaa atggtgccct
     1501 cccgataagt tggtgaacct gcacgacttt cgctcagacg aaattgagca ccttgttgta
     1561 gaggagttgg tcaggaagag agaggagtgt ctggatgcac tagagtccat catgacaacc
     1621 aagtcagtga gtttcagacg tctcagtcat ttaagaaaac ttgtccctgg gtttggaaaa
     1681 gcatatacca tattcaacaa gaccttgatg gaagccgatg ctcactacaa gtcagtcaga
     1741 acttggaatg agatcctccc ttcaaaaggg tgtttaagag ttggggggag gtgtcatcct
     1801 catgtgaacg gggtgttttt caatggtata atattaggac ctgacggcaa tgtcttaatc
     1861 ccagagatgc aatcatccct cctccagcaa catatggagt tgttggaatc ctcggttatc
     1921 ccccttgtgc accccctggc agacccgtct accgttttca aggacggtga cgaggctgag
     1981 gattttgttg aagttcacct tcccgatgtg cacaatcagg tctcaggagt tgacttgggt
     2041 ctcccgaact gggggaagta tgtattactg agtgcagggg ccctgactgc cttgatgttg
     2101 ataattttcc tgatgacatg ttgtagaaga gtcaatcgat cagaacctac gcaacacaat
     2161 ctcagaggga cagggaggga ggtgtcagtc actccccaaa gcgggaagat catatcttca
     2221 tgggaatcac acaagagtgg gggtgagacc agactgtaag cggccgctcg agcatgcatc
     2281 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2341 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2401 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2461 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2521 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2581 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2641 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     2701 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     2761 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     2821 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     2881 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     2941 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3001 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3061 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3121 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3181 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3241 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3301 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3361 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3421 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3481 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3541 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3601 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3661 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     3721 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     3781 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     3841 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     3901 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     3961 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4021 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4081 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4141 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4201 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4261 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4321 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4381 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4441 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4501 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4561 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4621 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4681 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4741 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4801 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4861 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4921 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4981 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5041 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5101 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5161 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5221 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5281 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5341 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5401 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5461 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5521 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5581 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5641 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5701 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5761 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5821 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5881 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5941 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6001 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6061 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6121 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6181 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6241 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6301 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6361 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6421 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6481 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6541 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6601 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6661 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6721 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     6781 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     6841 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     6901 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     6961 ccagatatac gcgtt
//
