LOCUS       #36.pA.NGFR.mCMV        9285 bp DNA     circular SYN 07-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9285)
  AUTHORS   Ferdinando Pucci
  TITLE     Direct Submission
  JOURNAL   Exported Sep 9, 2024 from SnapGene Viewer 7.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9285
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     attenuator      80..285
                     /label=40polyA oriR
     CDS             286..446
                     /label=C19
     misc_feature    447..905
                     /label=ageF1 origin
     misc_feature    906..2904
                     /label=C19
     primer_bind     3055..3075
                     /label=p1074 F primer 1 3050-3550
     attenuator      3122..3794
                     /label=V IE-I prom
     primer_bind     3412..3432
                     /label=p1074 F primer 2 3450-3950
     primer_bind     3726..3746
                     /label=p1074 F primer 3 3850-4350
     enhancer        3796..3977
                     /label=5
     attenuator      3978..4128
                     /label=S SL123
     misc_feature    4129..4483
                     /label=4 mgag
     primer_bind     4242..4262
                     /label=p1074 F primer 4 4250-4750
     primer_bind     4646..4666
                     /label=p1074 F primer 5 4250-4750
     primer_bind     4964..4984
                     /label=p1074 F primer 6 5050-5550
     primer_bind     5339..5359
                     /label=p1074 F primer 7 5450-5950
     misc_feature    complement(5502..5757)
                     /label=40 polyA unidirectional
     primer_bind     5812..5832
                     /label=p1074 F primer 8 5850-6350
     CDS             complement(5843..6685)
                     /codon_start=1
                     /label=NGFR
                     /translation="MGAGATGRAMDGPRLLLLLLLGVSLGGAKEACPTGLYTHSGECCK
                     ACNLGEGVAQPCGANQTVCEPCLDSVTFSDVVSATEPCKPCTECVGLQSMSAPCVEADD
                     AVCRCAYGYYQDETTGRCEACRVCEAGSGLVFSCQDKQNTVCEECPDGTYSDEANHVDP
                     CLPCTVCEDTERQLRECTRWADAECEEIPGRWITRSTPPEGSDSTAPSTQEPEAPPEQD
                     LIASTVAGVVTTVMGSSQPVVTRGTTDNLIPVYCSILAAVVVGLVAYIAFKRWNRGIL"
     misc_structure  5870..5935
                     /label=TM
     primer_bind     6097..6117
                     /label=p1074 F primer 9 6250-6750
     primer_bind     6543..6563
                     /label=p1074 F primer 10 6650-7150
     attenuator      6816..6924
                     /label=nhCMV
     primer_bind     6932..6952
                     /label=p1074 F primer 11 7050-7550
     enhancer        6958..7472
                     /label=PGK
     primer_bind     7443..7463
                     /label=p1074 F primer 12 7450-7950
     regulatory      7487..7495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     7493..7543
                     /codon_start=1
                     /label=SS
                     /translation="MGVKVLFALICIAVAEA"
     CDS             7544..7567
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             7583..8086
                     /codon_start=1
                     /label=GLuc
                     /translation="KPTENNEDFNIVAVASNFATTDLDADRGKLPGKKLPLEVLKEMEA
                     NARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESAQGGIGEAIVDIPEIPGF
                     KDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQRCATFASKIQGQVDKIK
                     GAGGD"
     CDS             8141..8293
                     /codon_start=1
                     /label=PDGFR TM
                     /translation="NAVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLW
                     QKKPR"
     primer_bind     8294..8313
                     /label=p1074 F primer 14 8250-8750
     misc_feature    8300..8806
                     /label=WPRE*
                     /note="6bp downstream SacII-site removed by linker insert 
                     (partial homology retained though)"
     primer_bind     8657..8677
                     /label=p1074 F primer 15 8650-9150
     primer_bind     8887..8907
                     /label=p1074 F primer 16 9050-9550
     enhancer        8931..9165
                     /label=3RU5
ORIGIN
        1 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
       61 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc cagttccgcc
      121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
      181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcgtc
      241 gagacgtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
      301 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
      361 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
      421 tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg
      481 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      541 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      601 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      661 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
      721 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
      781 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      841 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
      901 tttcccaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
      961 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1021 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1081 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1141 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1201 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1261 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1321 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1381 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1441 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1501 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1561 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     1621 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1681 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1741 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1801 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     1861 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1921 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1981 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2041 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2101 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2161 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2221 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2281 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2341 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2401 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2461 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2521 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2581 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2641 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2701 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2761 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     2821 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     2881 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     2941 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     3001 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     3061 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agctgcaagc
     3121 ttggccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
     3181 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     3241 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     3301 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     3361 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     3421 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     3481 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     3541 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     3601 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     3661 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     3721 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     3781 tcgtttagtg aaccggggtc tctctggtta gaccagatct gagcctggga gctctctggc
     3841 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     3901 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     3961 tggaaaatct ctagcagtgg cgcccgaaca gggacctgaa agcgaaaggg aaaccagagc
     4021 tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag gggcggcgac
     4081 tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat gggtgcgaga
     4141 gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg ttaaggccag
     4201 ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag ctagaacgat
     4261 tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata ctgggacagc
     4321 tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat acagtagcaa
     4381 ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct ttagacaaga
     4441 tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct gatcttcaga
     4501 cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata taaagtagta
     4561 aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt gcagagagaa
     4621 aaaagagcag tgggaatagg agctttgttc cttgggttct tgggagcagc aggaagcact
     4681 atgggcgcag cgtcaatgac gctgacggta caggccagac aattattgtc tggtatagtg
     4741 cagcagcaga acaatttgct gagggctatt gaggcgcaac agcatctgtt gcaactcaca
     4801 gtctggggca tcaagcagct ccaggcaaga atcctggctg tggaaagata cctaaaggat
     4861 caacagctcc tggggatttg gggttgctct ggaaaactca tttgcaccac tgctgtgcct
     4921 tggaatgcta gttggagtaa taaatctctg gaacagattt ggaatcacac gacctggatg
     4981 gagtgggaca gagaaattaa caattacaca agcttaatac actccttaat tgaagaatcg
     5041 caaaaccagc aagaaaagaa tgaacaagaa ttattggaat tagataaatg ggcaagtttg
     5101 tggaattggt ttaacataac aaattggctg tggtatataa aattattcat aatgatagta
     5161 ggaggcttgg taggtttaag aatagttttt gctgtacttt ctatagtgaa tagagttagg
     5221 cagggatatt caccattatc gtttcagacc cacctcccaa ccccgagggg acccgacagg
     5281 cccgaaggaa tagaagaaga aggtggagag agagacagag acagatccat tcgattagtg
     5341 aacggatctc gacggtatcg gttaactttt aaaagaaaag gggggattgg ggggtacagt
     5401 gcaggggaaa gaatagtaga cataatagca acagacatac aaactaaaga attacaaaaa
     5461 caaattacaa aaattcaaaa ttttatcgat cacgagacta gcctcgagag atctgatcat
     5521 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5581 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5641 taatggttac aaataaggca atagcatcac aaatttcaca aataaggcat ttttttcact
     5701 gcattctagt tttggtttgt ccaaactcat caatgtatct tatcatgtct ggatctcaaa
     5761 tccctcggaa gctgcgcctg tcatcgaatt cctgcagccc ggtgcatgac taagctagct
     5821 cagttagcct cccccatctc ccctagagga tccccctgtt ccacctcttg aaggctatgt
     5881 aggccacaag gcccacaacc acagcagcca ggatggagca atagacaggg atgaggttgt
     5941 cggtggtgcc tcgggtcacc acgggctggg agctgcccat cactgtggtc accacacctg
     6001 ccaccgtgct ggctatgagg tcttgttctg gaggtgcctc aggctcctgg gtgctggggg
     6061 ctgtgctgtc cgagccctct gggggtgtgg accgtgtaat ccaacggcca gggatctcct
     6121 cgcactcggc gtcggcccag cgtgtgcact cgcggagctg gcgctcggtg tcctcgcaca
     6181 cggtgcaggg caggcacggg tccacgtggt tggcctcgtc ggaatacgtg ccgtcggggc
     6241 actcctcgca cacggtgttc tgcttgtcct ggcaggagaa cacgaggccc gagcccgcct
     6301 cgcacacgcg gcacgcctcg cagcgcccag tcgtctcatc ctggtagtag ccgtaggcgc
     6361 agcggcacac ggcgtcgtcg gcctccacgc acggcgccga catgctctgg agccccacgc
     6421 actcggtgca cggcttgcac ggctcggtcg cgctcaccac gtcggagaac gtcacgctgt
     6481 ccaggcaggg ctcacacacg gtctggttgg ctccacaagg ctgggccaca ccctcgccca
     6541 ggttgcaggc tttgcagcac tcaccgctgt gtgtgtacag gcctgtgggg catgcctcct
     6601 tggcacctcc aagggacacc cccagaagca gcaacagcag caggcgcggc ccgtccatgg
     6661 cgcggccggt ggcacctgcc cccatcgccc gcctcccgcg gcagcgctcg acttccagct
     6721 cggtccgctt tgcggactga tggggctgcg ctgcgctgcg ctccagcgcc ccccctgccc
     6781 gccggagctg gccgcggccc gaattccgcg gaggctggat cggtcccggt gtcttctatg
     6841 gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct
     6901 gcttatatag gcctcccacc gtacacgcct accctcgaga agcttgatat cgaattccac
     6961 ggggttgggg ttgcgccttt tccaaggcag ccctgggttt gcgcagggac gcggctgctc
     7021 tgggcgtggt tccgggaaac gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc
     7081 cgttcgcagc gtcacccgga tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc
     7141 tgctccgccc ctaagtcggg aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac
     7201 ggaagccgca cgtctcacta gtaccctcgc agacggacag cgccagggag caatggcagc
     7261 gcgccgaccg cgatgggctg tggccaatag cggctgctca gcagggcgcg ccgagagcag
     7321 cggccgggaa ggggcggtgc gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct
     7381 gcccgcgcgg tgttccgcat tctgcaagcc tccggagcgc acgtcggcag tcggctccct
     7441 cgttgaccga atcaccgacc tctctcccca gggggatcca ccggtcgcca ccatgggcgt
     7501 aaaagtgctt tttgctctca tctgtattgc cgtggcagag gctgactaca aggacgacga
     7561 cgataagggg agtggactcg aaaaaccaac agagaataac gaggacttta atatagtggc
     7621 agtggcctca aatttcgcca caactgatct ggatgcagat aggggaaaac tccctgggaa
     7681 gaaactccca ctggaagtgt tgaaagaaat ggaggctaac gcccgaaagg ctgggtgtac
     7741 ccgaggttgt cttatttgtc tcagtcatat aaagtgtaca ccaaagatga agaaattcat
     7801 acccggacgc tgccatacct acgagggaga caaagaatcc gctcaggggg ggatagggga
     7861 ggcaatcgtg gacatccctg agattccagg attcaaagac cttgaaccca tggagcaatt
     7921 catcgcacag gttgatctct gcgtagattg tactacaggc tgcctgaagg gattggccaa
     7981 tgtccagtgt agcgacctgc tcaaaaaatg gctcccccaa aggtgtgcaa ctttcgcctc
     8041 aaaaatccaa ggccaggttg ataagattaa gggtgctggc ggagacggct ccaccagtgg
     8101 ttcaggtaag cccggatctg gagaaggctc cactaaaggg aacgcagtcg gccaagatac
     8161 acaggaggta attgtagtac ctcatagcct tcccttcaaa gtcgtggtaa tatccgcaat
     8221 actggcactg gtcgtcctta ctataatttc acttataata ctcattatgc tttggcaaaa
     8281 gaagccacgc taggtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg
     8341 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     8401 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     8461 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     8521 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     8581 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     8641 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac
     8701 gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     8761 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     8821 agagcctctt ccgcgtcttc gccttcccgg gtcgagctcg gtacctttaa gaccaatgac
     8881 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     8941 aattcactcc caacgaagac aagatctgct ttttgcttgt actgggtctc tctggttaga
     9001 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     9061 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     9121 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     9181 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     9241 gtttattgca gcttataatg gttacaaata aagcaatagc atcac
//
