LOCUS       194691                  6647 bp    DNA     circular UNA 26-JUN-2023
DEFINITION  CaProLa_PosCtrl-cpHalo_replaces_cpHaloDel.
ACCESSION   urn.local...7y-7vqwj6x
VERSION     urn.local...7y-7vqwj6x
KEYWORDS    .
SOURCE      mamalian
  ORGANISM  mamalian.
FEATURES             Location/Qualifiers
     misc_feature    16..604
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /standard_name="WPRE"
     primer_bind     complement(69..89)
                     /note="WPRE, reverse primer"
                     /standard_name="WPRE-R"
     primer_bind     complement(74..90)
                     /standard_name="WRPE.r"
     CDS             complement(487..498)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
                     /standard_name="Factor Xa site"
     primer_bind     complement(607..623)
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /standard_name="KS primer"
     primer_bind     complement(608..624)
                     /note="For pBluescript vector"
                     /standard_name="pBluescriptKS"
     polyA_signal    653..774
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(664..683)
                     /note="SV40 polyA terminator, reverse primer"
                     /standard_name="EBV-rev"
     primer_bind     718..737
                     /note="SV40 polyA, reverse primer"
                     /standard_name="SV40pA-R"
     repeat_region   936..1065
                     /standard_name="AAV2 ITR"
     primer_bind     complement(1131..1148)
                     /note="L4440 vector, forward primer"
                     /standard_name="L4440"
     rep_origin      complement(1302..1890)
     primer_bind     complement(1379..1396)
                     /standard_name="aavseq5"
     primer_bind     complement(1382..1401)
                     /note="pBR322 origin, forward primer"
                     /standard_name="pBR322ori-F"
     CDS             complement(2061..2921)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /standard_name="AmpR"
     primer_bind     2684..2703
                     /note="Ampicillin resistance gene, reverse primer"
                     /standard_name="Amp-R"
     promoter        complement(2922..3026)
                     /gene="bla"
                     /standard_name="AmpR promoter"
     rep_origin      complement(3052..3507)
     primer_bind     complement(3189..3210)
                     /note="F1 origin, forward primer"
                     /standard_name="F1ori-F"
     primer_bind     3401..3420
                     /note="F1 origin, reverse primer"
                     /standard_name="F1ori-R"
     primer_bind     3401..3414
                     /standard_name="paavseq3"
     repeat_region   3579..3708
                     /standard_name="AAV2 ITR"
     promoter        3790..4237
                     /note="human synapsin I promoter; confers neuron-specific
                     expression (Kugler et al., 2003)"
                     /standard_name="hSyn promoter"
     primer_bind     complement(3790..3804)
                     /standard_name="paavseq4"
     primer_bind     4208..4220
                     /standard_name="hSyn_new"
     regulatory      4269..>4274
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation
                      of translation (Kozak, 1987)"
     CDS             4275..4337
                     /standard_name="NES (CaMPARI)"
     misc_feature    4338..4340
                     /note="Geneious type: motif"
                     /standard_name="A145"
     CDS             <4338..>4367
                     /standard_name="HaloPep145-154"
     misc_feature    4362..4364
                     /note="Geneious type: motif"
                     /standard_name="R153"
     misc_feature    4365..4367
                     /note="Geneious type: motif"
                     /standard_name="T154"
     misc_feature    4368..4373
                     /note="Geneious type: motif"
                     /standard_name="GS"
     CDS             4374..4814
                     /codon_start=1
                     /product="calcium-binding protein"
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTE
                     AELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVVMMTAK"
     CDS             4821..4910
                     /standard_name="Pro30"
     misc_feature    4911..4919
                     /note="Geneious type: motif"
                     /standard_name="GGS"
     CDS             4920..4997
                     /standard_name="M13_2.07"
     misc_feature    4998..5006
                     /note="Geneious type: motif"
                     /standard_name="GGS"
     misc_feature    5007..5009
                     /note="Geneious type: motif"
                     /standard_name="F144"
     CDS             <5007..>5462
                     /note="Geneious type: cds"
                     /standard_name="HaloTag7"
     misc_feature    5463..5507
                     /note="Geneious type: motif"
                     /standard_name="(GGS/T)x5"
     CDS             <5508..>5921
                     /note="Geneious type: cds"
                     /standard_name="HaloTag7"
     misc_feature    5919..5921
                     /note="Geneious type: motif"
                     /standard_name="Trp141"
     CDS             <5922..5930
                     /codon_start=1
                     /standard_name="GGS-Linker"
     CDS             5931..6647
                     /standard_name="mEGFP"
ORIGIN      
        1 tgaaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
       61 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
      121 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
      181 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
      241 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
      301 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
      361 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
      421 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
      481 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
      541 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
      601 ccgcatcgat accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac
      661 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
      721 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
      781 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
      841 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
      901 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
      961 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     1021 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ktrwkraatm
     1081 ssscammgmk ssgkragagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1141 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1201 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1261 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1321 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1381 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1441 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1501 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1561 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1621 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1681 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1741 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1801 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1861 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1921 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1981 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2041 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2101 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2161 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2221 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2281 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2341 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2401 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2461 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2521 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2581 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2641 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2701 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2761 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2821 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2881 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2941 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3001 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     3061 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3121 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3181 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3241 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3301 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3361 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3421 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3481 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     3541 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg
     3601 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     3661 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
     3721 ttaacccgcc atgctactta tctacgtagc catgctctag gaagatctct gcagagggcc
     3781 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
     3841 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
     3901 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
     3961 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
     4021 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
     4081 tcgcgtccgc gccgccgccg gcccagccgg acnncaccac gcgaggcgcg agataggggg
     4141 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
     4201 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagtcg aattcaagct gctagcaagg
     4261 atccacccgc caccatgctg cagaacgagc ttgctcttaa gttggctgga cttgatatta
     4321 acaagactgg aggttctgcc cgcgagacct tccaggcctt ccgcaccggt tctgaccaac
     4381 tgactgaaga gcagatcgca gaatttaaag aggctttctc cctatttgac aaggacgggg
     4441 atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg cagaacccca
     4501 cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacaatcg
     4561 acttccctga gttcctgaca atgatggcaa gaaaaatgaa agacacagac agtgaagaag
     4621 aaattagaga agcgttccgt gtgtttgata aggatggcaa tggctacatc agtgcagcag
     4681 agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag gttgatgaaa
     4741 tgatcaggga agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtagtga
     4801 tgatgacagc gaaggaattc cctcctcctc cgccaccgcc accaccgccg ccgccgccac
     4861 caccacctcc tccgcctcct ccaccaccac cgccaccgcc tccaccacct ggcggcagca
     4921 gggtcgactc atcacgtcgt aagttcaata agacaggtca cgcactgaga gctataggtc
     4981 ggctgagctc actcgagggc ggttcttttg cccgcgagac cttccaggcc ttccgcacca
     5041 ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga
     5101 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     5161 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     5221 cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc
     5281 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc
     5341 tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc
     5401 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga
     5461 ttggtggcac cggtggctct ggcggtaccg gtggctctgg cggttctatc ggtactggct
     5521 ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg
     5581 gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt
     5641 ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg
     5701 gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca
     5761 tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg
     5821 gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat
     5881 ttatggagtt catccgccct atcccgacct gggacgaatg gggaggttct atggtgagca
     5941 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     6001 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     6061 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     6121 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     6181 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     6241 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     6301 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     6361 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     6421 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     6481 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     6541 cccagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     6601 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaag
//
