LOCUS       Exported                7639 bp ds-DNA     circular SYN 24-FEB-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7639)
  AUTHORS   Justin Lab
  TITLE     Direct Submission
  JOURNAL   Exported Feb 10, 2022 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7639
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     primer_bind     138..175
                     /label=Fragment.FOR
                     /note="color: black; sequence: 
                     TCCTGCGGCCGCACGCGTACTTGTGGACTAAGTTTGTT"
     misc_feature    157..519
                     /label=CamKIIa(0.4)
                     /note="color: #a6acb3"
     primer_bind     complement(500..537)
                     /label=Fragment.REV
                     /note="color: black; sequence: 
                     ACCGGTTTCGAAGGATCCGCTGCCCCCAGAACTAGGGG"
     CDS             549..2042
                     /codon_start=1
                     /label=PHR
                     /note="color: #00ffff"
                     /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ
                     FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY
                     DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI
                     ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN
                     EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE
                     ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL
                     VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY
                     ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA
                     SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA"
     primer_bind     complement(1697..1719)
                     /label=PHR-D387A-R
                     /note="color: black; sequence: CCAAGTCGGCGGCGAGAAGGGTA"
     primer_bind     1697..1719
                     /label=PHR-D387A-F
                     /note="color: black; sequence: TACCCTTCTCGCCGCCGACTTGG"
     primer_bind     1971..1986
                     /label=PHR-CF
                     /note="color: black; sequence: GATACAGCACGCGAGC"
     primer_bind     2029..2055
                     /label=CRY2PHR-InsMu-F
                     /note="color: black; sequence: TGATCGGAGCAGCGGgCAGCGGCgcca"
     CDS             2052..2108
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /note="color: #cc99b2"
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     2075..2134
                     /label=Fragment 2.FOR
                     /note="color: black; sequence: 
                     gcaagcaggagacgtggaagaaaaccccggtcctATGAATGGAGCTATAGGAGGTGACC
                     T"
     CDS             2109..2618
                     /codon_start=1
                     /label=CIBN
                     /note="color: #993366"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKAAKFDTETKDCN
                     EAAKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     primer_bind     2632..2650
                     /label=Primer 1
                     /note="color: black; sequence: 
                     GAAAAGCTAGCCACCCCACCATGGTGAGCAAGGG"
     CDS             2637..3344
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2792..2803
                     /label=Primer 1
                     /note="color: black; sequence: 
                     GAAAAGCTAGCCACCCCACCATGGTGAGCAAGGG"
     misc_feature    3354..3367
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     CDS             3375..3722
                     /codon_start=1
                     /label=VAMP2
                     /note="color: #3366ff"
                     /translation="MSATAATVPPAAPAGEGGPPAPPPNLTSNRRLQQTQAQVDEVVDI
                     MRVNVDKVLERDQKLSELDDRADALQAGASQFETSAAKLKRKYWWKNLKMMIILGVICA
                     IILIIIIVYFSS"
     primer_bind     complement(3690..3734)
                     /label=Fragment 2.REV
                     /note="color: black; sequence: 
                     GAAGCTTGAGCTCGAGGGCGCGCCTTAAGAGCTgaagtaaactatgatgatgatgagga
                     t"
     misc_feature    3766..4354
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(4237..4248)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 4236"
                     /translation="IEGR"
     polyA_signal    4386..4862
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     repeat_region   4902..5042
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4"
     rep_origin      5117..5572
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        5854..5958
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             5959..6819
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 5959 .. 6027 / #ccffcc / signal sequence
                        2: 6028 .. 6819 / #ccffcc"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6990..7578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtacttg tggactaagt ttgttcacat
      181 ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact tgctcctggg
      241 cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc aggttttaaa
      301 gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca gggcgtttcg
      361 gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt gagcagccac
      421 agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt tgcactcagg
      481 agcacgggca ggcgagtggc ccctagttct gggggcagcg gatccttcga aaccggtgct
      541 agcccaccat gaagatggac aaaaagacca tcgtctggtt tcggagagat ttgagaatag
      601 aagataatcc cgcgctcgcc gccgcggccc acgagggttc cgtcttcccc gttttcattt
      661 ggtgtcctga agaagaaggc cagttttatc ccggaagggc ctctaggtgg tggatgaagc
      721 aaagtctggc ccatcttagc cagtcactga aagcactggg cagtgatctt accctgatca
      781 agacacacaa taccatctct gccatcctcg actgcatcag ggtgaccggc gcaacgaaag
      841 tcgtgtttaa ccacctgtac gatccagtta gtctggtgcg cgaccacact gtgaaggaga
      901 agctggtgga acgggggatc agtgtgcaga gctacaacgg ggaccttctg tacgagccat
      961 gggagatcta ttgcgagaaa gggaaaccgt tcacctcctt caacagttac tggaagaaat
     1021 gtttggatat gtcaatagag tccgttatgt tgccccctcc ctggagactg atgccgatta
     1081 ctgctgctgc agaggccatc tgggcctgct ccatcgagga actcggtctg gaaaatgaag
     1141 cagaaaagcc aagcaatgca cttctcacta gagcctggag ccccggctgg tctaatgccg
     1201 acaagctgct taacgagttc atcgaaaaac aactgattga ctacgcgaag aactccaaga
     1261 aagtggtagg taactcaact agcttgctct ctccatatct ccattttggc gagatttctg
     1321 tccgccatgt atttcagtgc gctcggatga aacagattat ctgggctcgc gataaaaaca
     1381 gcgaaggcga agaaagcgcc gatctgttcc tgcgagggat cggacttcgg gaatactccc
     1441 ggtatatatg tttcaacttt ccattcacac acgagcagag tctgttgtcc cacctcaggt
     1501 tcttcccctg ggacgccgat gtcgacaaat tcaaggcatg gagacaggga aggacagggt
     1561 acccactcgt ggatgctggc atgagagagc tctgggctac aggctggatg cacaaccgca
     1621 tccgggtaat cgtgtcctca tttgctgtca agtttctgct cctgccttgg aaatggggaa
     1681 tgaagtactt ttgggatacc cttctcgacg ccgacttgga gtgtgacatt ctgggatggc
     1741 aatatattag cgggtcaatt cctgacggcc atgagttgga caggttggac aatccggcct
     1801 tgcagggagc taagtatgat cccgaaggag agtatattcg acagtggctc cccgagctgg
     1861 cccgacttcc tacggagtgg attcaccatc cttgggacgc accactgaca gtgctcaagg
     1921 caagcggggt ggagctgggc accaattacg ctaagcctat agttgatata gatacagcac
     1981 gcgagctgct ggctaaagcg atctctcgca ctcgggaggc gcagattatg atcggtgctg
     2041 ccggcagcgg cgccacgaac ttctctctgt taaagcaagc aggagacgtg gaagaaaacc
     2101 ccggtcctat gaatggagct ataggaggtg accttttgct caattttcct gacatgtcgg
     2161 tcctagagcg ccaaagggct cacctcaagt acctcaatcc cacctttgat tctcctctcg
     2221 ccggcttctt tgccgattct tcaatgatta ccggcggcga gatggacagc tatctttcga
     2281 ctgccggttt gaatcttccg atgatgtacg gtgagacgac ggtggaaggt gattcaagac
     2341 tctcaatttc gccggaaacg acgcttggga ctggaaattt caaggcagcg aagtttgata
     2401 cagagactaa ggattgtaat gaggcggcga agaagatgac gatgaacaga gatgacctag
     2461 tagaagaagg agaagaagag aagtcgaaaa taacagagca aaacaatggg agcacaaaaa
     2521 gcatcaagaa gatgaaacac aaagccaaga aagaagagaa caatttctct aatgattcat
     2581 ctaaagtgac gaaggaattg gagaaaacgg attatattca tgtaccggtc gccaccatgg
     2641 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     2701 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2761 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     2821 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     2881 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     2941 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     3001 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     3061 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     3121 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     3181 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     3241 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
     3301 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggcagt ggttccggac
     3361 tcagatctcg agctatgtcg gctaccgctg ccaccgtccc gcctgccgcc ccggccggcg
     3421 agggtggccc ccctgcacct cctccaaacc ttactagtaa caggagactg cagcagaccc
     3481 aggcccaggt ggatgaggtg gtggacatca tgagggtgaa tgtggacaag gtcctggagc
     3541 gggaccagaa gttgtcggag ctggatgacc gtgcagatgc cctccaggca ggggcctccc
     3601 agtttgaaac aagtgcagcc aagctcaagc gcaaatactg gtggaaaaac ctcaagatga
     3661 tgatcatctt gggagtgatc tgcgccatca tcctcatcat catcatagtt tacttcagct
     3721 cttaaggcgc gcccctgcag ggaattcgat atcaagctta tcgataatca acctctggat
     3781 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3841 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3901 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3961 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     4021 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     4081 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     4141 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta tgttgccacc
     4201 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     4261 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     4321 acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga
     4381 tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca
     4441 ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg
     4501 tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga
     4561 agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat
     4621 cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg
     4681 agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga
     4741 gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc
     4801 caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc
     4861 ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     4921 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4981 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     5041 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     5101 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5161 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5221 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     5281 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     5341 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5401 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc
     5461 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     5521 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     5581 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     5641 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5701 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5761 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5821 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5881 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5941 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     6001 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     6061 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     6121 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     6181 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     6241 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     6301 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     6361 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     6421 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6481 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6541 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6601 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6661 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6721 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6781 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6841 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6901 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6961 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     7021 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     7081 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     7141 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     7201 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     7261 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     7321 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     7381 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     7441 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7501 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7561 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7621 tggccttttg ctcacatgt
//
