LOCUS       pAAV-Neo_CAG        7494 bp DNA     circular SYN 30-APR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7494)
  AUTHORS   A Bertero
  TITLE     Direct Submission
  JOURNAL   Exported Apr 30, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    402..1202
                     /label=AAVS1 HA-L
     misc_feature    1210..1236
                     /label=SA
     CDS             join(1237..1240,1241..1251,1252..1260)
                     /codon_start=1
                     /label=linker
                     /translation="GLERSGSG"
     CDS             1261..1314
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1324..2127
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    2165..2351
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        2451..2754
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2756..3033
                     /label=chicken beta-actin promoter
     intron          3035..4052
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     polyA_signal    4311..4392
                     /label=SV40 pA
                     /note="SV40 polyadenylation signal"
     misc_feature    4418..5254
                     /label=AAVS1 HA-R
                     /note="right homology arm of AAVS1"
     rep_origin      complement(5675..6263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6434..7294)
                     /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"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtgctttctc tgaccagcat
      421 tctctcccct gggcctgtgc cgctttctgt ctgtagcttg tggcctgggt cacctctacg
      481 gctggcccag atccttccct gccgcctcct tcaggttccg tcttcctcca ctccctcttc
      541 cccttgctct ctgctgtgtt gctgcccaag gatgctcttt ccggagcact tccttctcgg
      601 cgctgcacca cgtgatgtcc tctgagcgga tcctccccgt gtctgggtcc tctccgggca
      661 tctctcctcc ctcacccaac cccatgccgt cttcactcgc tgggttccct tttccttctc
      721 cttctggggc ctgtgccatc tctcgtttct taggatggcc ttctccgacg gatgtctccc
      781 ttgcgtcccg cctccccttc ttgtaggcct gcatcatcac cgtttttctg gacaacccca
      841 aagtaccccg tctccctggc tttagccacc tctccatcct cttgctttct ttgcctggac
      901 accccgttct cctgtggatt cgggtcacct ctcactcctt tcatttgggc agctccccta
      961 ccccccttac ctctctagtc tgtgctagct cttccagccc cctgtcatgg catcttccag
     1021 gggtccgaga gctcagctag tcttcttcct ccaacccggg cccctatgtc cacttcagga
     1081 cagcatgttt gctgcctcca gggatcctgt gtccccgagc tgggaccacc ttatattccc
     1141 agggccggtt aatgtggctc tggttctggg tacttttatc tgtcccctcc accccacagt
     1201 ggggcaagct tctgacctct tctcttcctc ccacagggcc tcgagagatc tggcagcgga
     1261 gagggcagag gaagtcttct aacatgcggt gacgtggagg agaatcccgg ccctaggctc
     1321 gagatgggat cggccattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     1381 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     1441 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     1501 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     1561 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     1621 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     1681 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     1741 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     1801 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     1861 cgcatgcccg acggcgatga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     1921 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     1981 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     2041 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     2101 tatcgccttc ttgacgagtt cttctgatct agagggcccg tttaaacccg ctgatcagcc
     2161 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     2221 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     2281 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     2341 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggt cacttattaa
     2401 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     2461 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     2521 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     2581 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     2641 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     2701 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     2761 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     2821 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
     2881 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     2941 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     3001 cggccctata aaaagcgaag cgcgcggcgg gcggggagtc gctgcgacgc tgccttcgcc
     3061 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     3121 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag cgcttggttt
     3181 aatgacggct tgtttctttt ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc
     3241 ctttgtgcgg ggggagcggc tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg
     3301 cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg
     3361 cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     3421 gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg
     3481 tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc tccccgagtt gctgagcacg
     3541 gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg
     3601 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     3661 cgggggaggg gcgcggcggc ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca
     3721 gccattgcct tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct
     3781 gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc
     3841 ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc
     3901 gtccccttct ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg
     3961 gacggggcag ggcggggttc ggcttctggc gtgtgaccgg cggctctaga gcctctgcta
     4021 accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg gttattgtgc
     4081 tgtctcatca ttttggcaaa gaattaattc ggatcaattc gagctcggta cccggggatc
     4141 ctctagtcag ctgacgcgtg ctagcgcggc cgcatcgata agcttgtcga cgatatctct
     4201 agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     4261 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     4321 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     4381 ttttttcact gccttgacag tactcttaag tcgacttact agggacagga ttggtgacag
     4441 aaaagcccca tccttaggcc tcctccttcc tagtctcctg atattgggtc taacccccac
     4501 ctcctgttag gcagattcct tatctggtga cacaccccca tttcctggag ccatctctct
     4561 ccttgccaga acctctaagg tttgcttacg atggagccag agaggatcct gggagggaga
     4621 gcttggcagg gggtgggagg gaaggggggg atgcgtgacc tgcccggttc tcagtggcca
     4681 ccctgcgcta ccctctccca gaacctgagc tgctctgacg cggctgtctg gtgcgtttca
     4741 ctgatcctgg tgctgcagct tccttacact tcccaagagg agaagcagtt tggaaaaaca
     4801 aaatcagaat aagttggtcc tgagttctaa ctttggctct tcacctttct agtccccaat
     4861 ttatattgtt cctccgtgcg tcagttttac ctgtgagata aggccagtag ccagccccgt
     4921 cctggcaggg ctgtggtgag gaggggggtg tccgtgtgga aaactccctt tgtgagaatg
     4981 gtgcgtccta ggtgttcacc aggtcgtggc cgcctctact ccctttctct ttctccatcc
     5041 ttctttcctt aaagagtccc cagtgctatc tgggacatat tcctccgccc agagcagggt
     5101 cccgcttccc taaggccctg ctctgggctt ctgggtttga gtccttggca agcccaggag
     5161 aggcgctcag gcttccctgt cccccttcct cgtccaccat ctcatgcccc tggctctcct
     5221 gccccttccc tacaggggtt cctggctctg ctctaagctt ggcgtaatca tggtcatagc
     5281 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5341 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5401 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5581 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5641 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5701 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5761 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5821 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5881 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5941 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6001 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6061 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6121 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6181 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6241 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6301 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6361 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6421 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6481 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6541 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6601 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6661 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6721 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6781 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6841 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6901 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6961 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7021 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7081 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7141 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7201 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7261 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7321 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7381 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7441 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
