LOCUS       Exported                7174 bp DNA     circular SYN 12-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7174)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 7, 2021 from SnapGene 5.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7174
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        163..699
                     /label=mDlx
     intron          860..992
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     975..994
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1046..1079
                     /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."
     protein_bind    1130..1163
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1185..2756)
                     /codon_start=1
                     /label=N2C-G
                     /translation="VPQVLLFVLLLGFSLCFGKFPIYTIPDELGPWSPIDIHHLSCPNN
                     LVVEDEGCTNLSEFSYMELKVGYISAIKVNGFTCTGVVTEAETYTNFVGYVTTTFKRKH
                     FRPTPDACRAAYNWKMAGDPRYEESLHNPYPDYHWLRTVRTTKESLIIISPSVTDLDPY
                     DKSLHSRVFPGGKCSGITVSSTYCSTNHDYTIWMPENPRPRTPCDIFTNSRGKRASNGN
                     KTCGFVDERGLYKSLKGACRLKLCGVLGLRLMDGTWVAMQTSDETKWCPPDQLVNLHDF
                     RSDEIEHLVVEELVKKREECLDALESIMTTKSVSFRRLSHLRKLVPGFGKAYTIFNKTL
                     MEADAHYKSVRTWNEIIPSKGCLKVGGRCHPHVNGVFFNGIILGPDDHVLIPEMQSSLL
                     QQHMELLESSVIPLMHPLADPSTVFKEGDEAEDFVEVHLPDVYKQISGVDLGLPNWGKY
                     VLMTAGAMIGLVLIFSLMTWCRRANRPESKQRSFGGTGGNVSVTSQSGKVIPSWESYKS
                     GGEIRL"
     CDS             complement(2757..2759)
                     /codon_start=1
                     /label=oG(B19)
                     /translation="M"
     CDS             complement(2763..2819)
                     /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."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     2822..2855
                     /label=Fragment.FOR
     primer_bind     2837..2923
                     /label=Primer 1
     primer_bind     complement(2839..2925)
                     /label=Primer 2
     CDS             complement(2841..2906)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2910..2940)
                     /label=Fragment.REV
     CDS             complement(2928..3365)
                     /codon_start=1
                     /label=TVA
                     /translation="MARLLPALLLLLLPGNVTGNGSGNGFLSRCPPGQFRCSEPPGAHG
                     ECYPQDWLCDGHPDCDDGRDEWGCGTSAIPAVPTDNGTEAPTAPASGRALPARNHGRMW
                     MLITAGPLCCLVAVGGIAAWGKSKAKSRFDIFSLERASKGGG"
     regulatory      3366..3371
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(3378..3411)
                     /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."
     protein_bind    complement(3462..3495)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3520..4108
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3587..3606)
                     /label=WHV5R
     primer_bind     3875..3887
                     /label=CCMV24
     CDS             complement(3991..4002)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4277..4358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4314..4333)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4368..4387
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   4437..4577
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4437..4566
                     /label=AAV2 ITR
     rep_origin      4652..5107
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4739..4758)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4949..4970
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5124..5143)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5243..5265
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5303..5321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5389..5493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5494..6354
                     /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     complement(5712..5731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6525..7113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7014..7033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaat taagaattct atacactcac
      181 agtggtttgg catatatttg gtgaaatttt ttaaggaaaa attagtgttg gtttcgatat
      241 atggtagctt tttctctaac ataatttgaa taattcagca aagccctact accagctgta
      301 cttctgcagc ctcttccatt ctttccagca ttataatttt ggttaatttt caattttagg
      361 tcctacgtct ctgcaatttg tgtatgaata acagaataat ttccctcttt tgtttcgcct
      421 ttcctgttcc tgaatctaaa taaagatggc tttttagtat taaaagtgga agaaaattac
      481 aggtaattat ctttgacggt aaaaacgctg taatcagcgg gctacatgaa aaattactct
      541 aattatggct gcatttaaga gaatggaaaa aaaccttctt gtggataaaa accttaaatt
      601 gtccccaatg tctgcttcaa attggatggc actgcagctg gaggctttgt tcagaattga
      661 tcctggggag ctacgaaccc aaagtttcac agtaggaagc ttgatatcga attcctgcag
      721 cccgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctagcct
      781 gcaggtcgag gagcgcagcc ttccagaagc agagcgcggc gccttaagct gcagaagttg
      841 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
      901 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
      961 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
     1021 ttaagaaact agagtcgact ccggaataac ttcgtatagg atactttata cgaagttatg
     1081 cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa taacttcgta
     1141 tagcatacat tatacgaagt tatggcgcgc cttacttgta catctcacag tctgatctca
     1201 cctccactct tatatgattc ccatgaaggt atgacttttc cgctttggga agtgactgac
     1261 acattccccc ctgtccctcc aaaactgcgt tgtttcgatt ctggtcgatt ggctcttctg
     1321 caccatgtca ttagggaaaa tatcaacacc aggccaatca tggcccctgc agtcatcaat
     1381 acatactttc cccagttcgg gagacccagg tcaacccctg agatctgttt gtacacatcg
     1441 gggaggtgaa cttcaacaaa atcctcagcc tcatcacctt ctttgaaaac tgtagaaggg
     1501 tctgccaggg ggtgcatcag ggggataact gaagattcca acaactccat atgttgctgg
     1561 aggagggatg attgcatctc tgggattagg acatggtcgt caggccctaa tattatacca
     1621 ttgaaaaaca ccccgttcac atgaggatgg caccttcctc caactttcaa acaccctttt
     1681 gaggggatga tctcattcca ggtccggact gacttgtagt gagcatcagc ctccatcaag
     1741 gttttgttga atatggtata tgcttttcca aaccctggga caagttttct caggtgactg
     1801 agacgtctga aacttactga cttggtggtc atgatggact ctaatgcatc cagacattcc
     1861 tctcttttct tgactaactc ctccacaacg agatgctcaa tctcgtctga gcgaaagtcg
     1921 tgcaaattca ccaactgatc tggagggcac catttggtct catctgatgt ttgcatcgcg
     1981 acccatgttc catccataag tctaagtcca agaactccac ataacttgag cctgcatgct
     2041 ccttttagag acttatacag gcctctttca tccacaaagc cgcaagtctt gttcccgttg
     2101 gatgctctct tccctctgct attggtaaaa atgtcacaag gtgtccttgg tctcggattc
     2161 tcgggcatcc aaatggtgta atcatggtta gttgagcagt aggtagagga caccgttatt
     2221 cctgagcact ttccgccagg gaagaccctt gagtgaaggg atttgtcata tgggtccaaa
     2281 tctgtcacac ttggggatat gataatgagg gactctttgg tggttcttac agttcgaagc
     2341 cagtggtagt cggggtatgg attgtgtagg gactcttcat atctggggtc accggccatc
     2401 ttccagttat acgcggctct acatgcgtct ggggtggggc ggaaatgctt tctcttgaat
     2461 gtggttgtga cataaccaac aaagttggtg taggtctctg cctctgtcac aacacctgtg
     2521 caagtgaacc cgttcacttt gatggctgag atgtatccca ctttgagttc catgtaggag
     2581 aactcggaca ggttggtaca tccttcatcc tccacaacca ggttatttgg acagctgaga
     2641 tggtgtatgt caatagggct ccagggacca agttcgtctg gtatcgtgta aatggggaac
     2701 ttcccgaaac acaacgaaaa acccagaagg agtacaaaca aaagaacctg aggaaccatg
     2761 gttggcccgg gattctcttc gacatcccct gcttgtttca acagggagaa gttagtggct
     2821 ccgcttccgt tgtcttcgaa cttgtcgtca tcgtctttgt agtcgatatc atgatcctta
     2881 taatcaccat catgatcctt ataatcgccg gtgctgcccg gttcgaaccc accccccttg
     2941 gatgcccttt caagactgaa aatgtcaaac ctgctttttg ctttggactt cccccatgca
     3001 gcgatacccc ccacagctac caggcagcac aggggccctg cagtgatcag catccacatg
     3061 cggccgtgat tcctggctgg caaagcacgt ccagaagcag gggcagtggg agcctctgtg
     3121 ccgttgtccg tgggcaccgc ggggatcgcg ctggtcccgc acccccactc gtcccgcccg
     3181 tcgtcgcagt cggggtgtcc gtcgcacagc cagtcctgcg ggtaacactc cccgtgggca
     3241 ccgggcggct ccgagcagcg gaactgaccg ggggggcaac gggacaaaaa accgttaccg
     3301 gacccgttac cggtcacgtt accgggcagc agcagcagca gcagcgcggg cagcagccgc
     3361 gccatggtgg cactagcata acttcgtata aagtatccta tacgaagtta tttgccttaa
     3421 cccagaaatt atcactgtta ttctttagaa tggtgcaaag aataacttcg tataatgtat
     3481 gctatacgaa gttatgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     3541 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3601 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3661 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3721 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3781 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3841 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3901 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     3961 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     4021 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     4081 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctgtg
     4141 atagcggcca tcaagctggc cgcgactcta gatcataatc agccatacca catttgtaga
     4201 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4261 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4321 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4381 actcatcaat gtatcagctt atcgataccg catgcacgtg cggaccgagc ggccgcagga
     4441 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4501 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4561 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4621 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4681 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4741 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4801 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4861 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4921 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4981 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     5041 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5101 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5161 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5221 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5281 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     5341 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5461 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5521 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5581 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5641 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5701 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5761 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5821 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5881 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5941 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6001 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6061 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6121 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6181 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6241 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6301 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6361 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6421 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6481 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6541 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6601 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6661 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6721 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6781 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6841 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6901 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6961 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7021 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7081 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7141 ttcctggcct tttgctggcc ttttgctcac atgt
//
