LOCUS       Exported                6254 bp ds-DNA     circular SYN 04-JUL-2024
DEFINITION  Express Rpl22HA in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6254)
  AUTHORS   Baljit Khakh
  TITLE     RiboTag AAV
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6254)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jul 5, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6254
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     128..145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    262..283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        298..328
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    336..352
                     /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)."
     primer_bind     341..363
                     /label=No.55 M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     360..376
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     360..376
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   414..543
                     /label=AAV2 ITR
     primer_bind     complement(425..436)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(426..441)
                     /label=No.37_Addgene-ITR_AAV2_Rv
     primer_bind     481..496
                     /label=No.37_Addgene-ITR_AAV2_Rv
     primer_bind     486..497
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(522..542)
                     /label=No.36_Addgene-ITR_AAV2_Fw
     primer_bind     523..543
                     /label=No.35_ITR-primer-Fw&Rv
     misc_feature    604..1284
                     /label=GfaABC1D promoter
     primer_bind     complement(949..960)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     1202..1222
                     /label=No56. GfaABC1D promotor
     intron          1394..1526
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1509..1528
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1571..1589
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1590..1606
                     /label=EGFP-5'insert
     primer_bind     1671..1685
                     /label=EGFP-5'insert
     protein_bind    complement(1698..1731)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     primer_bind     1742..1762
                     /label=No.47_5-loxp
     protein_bind    complement(1794..1827)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     1828..1868
                     /label=プライマー 2
     primer_bind     1836..1868
                     /label=No.52 mNeonGreen-Rv
     CDS             complement(1851..2561)
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      2297..2306
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     complement(2544..2587)
                     /label=プライマー 1
     primer_bind     complement(2556..2568)
                     /label=No.49 mScarlet-I-Fw
     regulatory      2558..2567
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    2584..2617
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     primer_bind     complement(2635..2655)
                     /label=No.48_3-lop
     protein_bind    2688..2721
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(2739..2768)
                     /label=No1.EGFP-3'insert
     misc_feature    2762..2774
                     /label=JN011-mini cloning site
     primer_bind     complement(2790..2822)
                     /label=No.50 mScarlet-I-Rv
     primer_bind     2808..2828
                     /label=No.41_WPRE-Fw-1
     misc_feature    2809..3416
                     /label=WPRE fron XCaMP-R
     misc_feature    2812..3404
                     /label=WPRE
                     /note="/vntifkey=21"
     primer_bind     complement(2823..2843)
                     /label=No24.WPRE-sec check
     primer_bind     complement(3399..3431)
                     /label=No.42_WPRE-Rv-1
     primer_bind     complement(3416..3431)
                     /label=No.43_WPRE-Rv-2
     polyA_signal    3434..3555
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3434..3451)
                     /label=No.43_WPRE-Rv-2
     primer_bind     complement(3445..3464)
                     /label=No.46_SV40-Rv
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3499..3518
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3717..3846
                     /label=AAV2 ITR
     primer_bind     complement(3717..3737)
                     /label=No.35_ITR-primer-Fw&Rv
     primer_bind     3718..3738
                     /label=No.36_Addgene-ITR_AAV2_Fw
     primer_bind     complement(3763..3774)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3764..3779)
                     /label=No.37_Addgene-ITR_AAV2_Rv
     primer_bind     3819..3834
                     /label=No.37_Addgene-ITR_AAV2_Rv
     primer_bind     3824..3835
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(3865..3882)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3865..3881)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3874..3896)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4023..4478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4110..4129)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4320..4341
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4504..4608
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4609..5469
                     /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"
     misc_feature    4812..4822
                     /label=1-sechceck-egfp
     primer_bind     complement(4827..4846)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5640..6228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6129..6148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
       61 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      121 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      181 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
      241 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      301 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      361 aggaaacagc tatgaccatg attacgccag atttaattaa ggccttaatt aggctgcgcg
      421 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
      481 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      541 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
      601 tctaacatat cctggtgtgg agtaggggac gctgctctga cagaggctcg ggggcctgag
      661 ctggctctgt gagctgggga ggaggcagac agccaggcct tgtctgcaag cagacctggc
      721 agcattgggc tggccgcccc ccagggcctc ctcttcatgc ccagtgaatg actcaccttg
      781 gcacagacac aatgttcggg gtgggcacag tgcctgcttc ccgccgcacc ccagcccccc
      841 tcaaatgcct tccgagaagc ccattgagca gggggcttgc attgcacccc agcctgacag
      901 cctggcatct tgggataaaa gcagcacagc cccctagggg ctgcccttgc tgtgtggcgc
      961 caccggcggt ggagaacaag gctctattca gcctgtgccc aggaaagggg atcaggggat
     1021 gcccaggcat ggacagtggg tggcaggggg ggagaggagg gctgtctgct tcccagaagt
     1081 ccaaggacac aaatgggtga ggggagagct ctccccatag ctgggctgcg gcccaacccc
     1141 accccctcag gctatgccag ggggtgttgc caggggcacc cgggcatcgc cagtctagcc
     1201 cactccttca taaagccctc gcatcccagg agcgagcaga gccagagcag gttggagagg
     1261 agacgcatca cctccgctgc tcgcaagctt tattgcggta gtttatcaca gttaaattgc
     1321 taacgcagtc agtgcttctg acacaacagt ctcgaactta agctgcagaa gttggtcgtg
     1381 aggcactggg caggtaagta tcaaggttac aagacaggtt taaggagacc aatagaaact
     1441 gggcttgtcg agacagagaa gactcttgcg tttctgatag gcacctattg gtcttactga
     1501 catccacttt gcctttctct ccacaggtgt ccactcccag ttcaattaca gctcttaagg
     1561 ctagagtact taatacgact cactataggc tagagcctct gctaaccatg ttcatgcctt
     1621 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     1681 caaagaattg gatccccata acttcgtata aagtatccta tacgaagtta tatcaaaata
     1741 ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca tcaccatcga cccataactt
     1801 cgtataatgt atgctatacg aagttatact agggatcccc cctcgagttt tcacttgtac
     1861 agctcgtcca tgcccatcac atcggtaaag gccttttgcc actccttgaa gttgagctcg
     1921 gtcttggagt gcttgagctc cgtcttacgg aacacgtaca tcggctggtt cttcagatag
     1981 ttagccgcca ttggcttggc aaaggtgtag gtggtccgcg cagtgctccg gtagcgcttg
     2041 ccatttccag tggtgtaact ccacttaaag gtactgatga tggttttgtc gttggggtaa
     2101 gtcttcttcg acctgcacca gtccgcagcg gtcagcgagt tggtcatcac aggaccgtca
     2161 gcagggaaac cagtcccctt cacctgggcc tctcctttga tgtggcttcc ctcgtaggtg
     2221 tagcggtagt taacagtaag ggaggcacca tcttcaaact gcattgtgcg atggacttgg
     2281 tatccggagc catctaccat ggcggcctgg aaaggcgaca tcccgtcagg gtagggcagg
     2341 tactgatgga agccataccc gatatgaggg accagaatcc agggggagaa ctggaggtca
     2401 cccttggtgg acttcaggtt taactcctca taaccatcat ttggattgcc ggtgccctga
     2461 cccaccatgt caaagtccac accgttgatg gagccaaaga tgtgtaactc atgtgtcgct
     2521 gggagagagg ccatgttatc ctcctcgccc ttgctcacca tggtggcggt accgaattct
     2581 agtataactt cgtataggat actttatacg aagttatcat tgggattctt cctattttga
     2641 tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccata acttcgtata
     2701 gcatacatta tacgaagtta tgtccctcga agaggttcga attcgatatc aagcttctgc
     2761 agtcgacgct agcgggccgc gactctagat cataatcagc cataccacgc ataatcaacc
     2821 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2881 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2941 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3001 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3061 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3121 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3181 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     3241 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3301 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3361 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcggccgg ccgcttgctt
     3421 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     3481 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     3541 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     3601 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     3661 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
     3721 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     3781 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     3841 gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     3901 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     3961 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4021 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4081 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4141 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4201 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4261 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4321 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     4381 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4441 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     4501 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4561 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4621 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4681 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4741 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4801 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4861 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4921 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4981 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5041 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5101 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5161 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     5221 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     5281 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     5341 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     5401 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     5461 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     5521 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5581 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5641 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5701 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5761 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5821 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5881 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5941 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6001 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6061 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6121 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     6181 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     6241 gcggcctttt tacg
//
