LOCUS       Exported                6285 bp ds-DNA     circular SYN 08-FEB-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6285)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Apr 13, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6285
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..249
                     /label=CMV enhancer
     enhancer        250..380
                     /label=CMV enhancer
     promoter        381..584
                     /label=CMV promoter
     misc_feature    510..534
                     /label=ACK15
     misc_feature    535..555
                     /label=CMV-F (genewiz)
     promoter        626..644
                     /label=T7 promoter
     misc_feature    complement(644..675)
                     /label=QC45*
     misc_feature    complement(644..660)
                     /label=QC43 anneal
     CDS             658..3543
                     /codon_start=1
                     /label=Translation 658-3543
                     /translation="MASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKV
                     TCSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCEL
                     IVKAMQGLLKDGNPIPSAIAANSGIYSSGGSSGGSSGSETPGTSESATPESSGGSSGGS
                     SSKTGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNK
                     HVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPNVTLFIYIARL
                     YHLANPRNRQGLRDLISSGVTIQIMTEQESGYCWHNFVNYSPSNESHWPRYPHLWVRLY
                     VLELYCIILGLPPCLNILRRKQSQLTSFTIALQSCHYQRLPPHILWATGLKSGGSSGGS
                     SGSETPGTSESATPESSGGSSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNK
                     PESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSGGSGGST
                     NLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSD
                     APEYKPWALVIQDSNGENKIKMLSGGSKRTADGSEFEPKKKRKVGSGATNFSLLKQAGD
                     VEENPGPGSGEGRGSLLTCGDVEENPGPMKRTADGSEFESPKKKRKVSSEVEFSHEYWM
                     RHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQ
                     NYRLYDATLYSTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHHPGMNHRV
                     EITEGILADECAALLCRFFRMPRRVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATP
                     ESSGGSSGGSKSIRCKNCNKLLFKADSFDHIEIRCPRCKRHIIMLNACEHPTEKHCGKR
                     EKITHSDETVRYGSKRTADGSEFEPKKKRKV"
     CDS             658..2526
                     /codon_start=1
                     /label=Translation 658-2526
                     /translation="MASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKV
                     TCSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCEL
                     IVKAMQGLLKDGNPIPSAIAANSGIYSSGGSSGGSSGSETPGTSESATPESSGGSSGGS
                     SSKTGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNK
                     HVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPNVTLFIYIARL
                     YHLANPRNRQGLRDLISSGVTIQIMTEQESGYCWHNFVNYSPSNESHWPRYPHLWVRLY
                     VLELYCIILGLPPCLNILRRKQSQLTSFTIALQSCHYQRLPPHILWATGLKSGGSSGGS
                     SGSETPGTSESATPESSGGSSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNK
                     PESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSGGSGGST
                     NLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSD
                     APEYKPWALVIQDSNGENKIKMLSGGSKRTADGSEFEPKKKRKVGSG"
     misc_feature    658..2526
                     /label=QTC161 (MCP-32-evo)
     misc_feature    658..681
                     /label=QC44*
     misc_feature    658..675
                     /label=USER MCP (1)
     misc_feature    661..1047
                     /label=MS2-CP (MCP)
     misc_feature    675
                     /label=silent A>T
     misc_feature    823..825
                     /label=N55K
     misc_feature    complement(1030..1050)
                     /label=QC45*
     misc_feature    1033..1069
                     /label=QC81
     misc_feature    1033..1050
                     /label=USER MCP (2)
     misc_feature    1050
                     /label=SILENT A>T
     misc_feature    1051..1146
                     /label=32 aa linker
     misc_feature    1051..1069
                     /label=ANNEAL
     misc_feature    complement(1133..1164)
                     /label=QC190
     misc_feature    1147..1926
                     /label=evo-rA1 frag
     misc_feature    1147..1830
                     /label=evo-rAPOBEC1
     misc_feature    1147..1168
                     /label=QC191
     misc_feature    1147..1164
                     /label=USER
     misc_feature    complement(1824..1842)
                     /label=195-anneal
     misc_feature    1831..1926
                     /label=32aa linker
     misc_feature    complement(1913..1926)
                     /label=a
     misc_feature    1917..1935
                     /label=194-anneal
     CDS             1927..2175
                     /label=UGI
     misc_feature    1927..1951
                     /label=QC50
     misc_feature    1927..1943
                     /label=USER UGI
     misc_feature    complement(1927..1943)
                     /label=QC49
     CDS             2206..2454
                     /label=UGI
     misc_feature    2467..2517
                     /label=bpNLS
     CDS             2497..2517
                     /label=SV40 NLS
     CDS             2527..2584
                     /label=P2A
     CDS             2527..2584
                     /codon_start=1
                     /label=Translation 2527-2584
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    2527..2551
                     /label=ACK2210
     misc_feature    complement(2565..2592)
                     /label=QC291 (R-BB/CBE)
     misc_feature    complement(2565..2583)
                     /label=anneal
     misc_feature    complement(2580..2632)
                     /label=Rev-T2A
     primer_bind     complement(2580..2632)
                     /label=R
     misc_feature    2580..2592
                     /label=USER
     CDS             2584..2646
                     /codon_start=1
                     /label=Translation 2584-2646
                     /translation="GSGEGRGSLLTCGDVEENPGP"
     CDS             2584..2646
                     /label=T2A
     misc_feature    2592
                     /label=silent mut
                     /label=silent mut.
     misc_feature    2593..2648
                     /label=Fwd-T2A
     primer_bind     2593..2648
                     /label=F
     misc_feature    2633..3516
                     /label=QTC135 (8.20)
     misc_feature    2633..2665
                     /label=QC288 (ABE-F)
     misc_feature    2633..2648
                     /label=USER
     CDS             2647..3543
                     /codon_start=1
                     /label=Translation 2647-3543
                     /translation="MKRTADGSEFESPKKKRKVSSEVEFSHEYWMRHALTLAKRARDER
                     EVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLYDATLYSTFE
                     PCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAAL
                     LCRFFRMPRRVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSKSIR
                     CKNCNKLLFKADSFDHIEIRCPRCKRHIIMLNACEHPTEKHCGKREKITHSDETVRYGS
                     KRTADGSEFEPKKKRKV"
     misc_feature    2650..2703
                     /label=bpNLS
     misc_feature    complement(2683..2719)
                     /label=QC209
     CDS             2683..2703
                     /label=SV40 NLS
     misc_feature    2704..2724
                     /label=QC 151
     misc_feature    2704..2719
                     /label=USER
     misc_feature    2707..3204
                     /label=TadA** (ABE8.20)
     misc_feature    complement(3187..3206)
                     /label=150anneal 1 mm
     misc_feature    3205..3300
                     /label=32aa linker
     misc_feature    complement(3283..3313)
                     /label=QC208
     misc_feature    complement(3283..3300)
                     /label=anneal
     misc_feature    3301..3483
                     /label=Com
     misc_feature    3301..3323
                     /label=QC113
     misc_feature    3301..3313
                     /label=USER
     misc_feature    complement(3466..3483)
                     /label=QC115
     misc_feature    3484..3486
                     /label=Linker
     misc_feature    3490..3540
                     /label=bpNLS
     misc_feature    complement(3492..3516)
                     /label=QC289
     misc_feature    complement(3492..3507)
                     /label=anneal ABE
     misc_feature    3498
                     /label=not in QTC135
     misc_feature    join(3503..6285,1..2592)
                     /label=QTC161 BB
     misc_feature    3503..3545
                     /label=QC290 (F-BB)
     misc_feature    3503..3516
                     /label=USER
     misc_feature    3508..3510
                     /label=new codon
     misc_feature    3513
                     /label=new codon
     misc_feature    3516
                     /label=new codon
     misc_feature    3519
                     /label=new codon
     misc_feature    3520..3545
                     /label=anneal BB
     CDS             3520..3540
                     /label=SV40 NLS
     CDS             3549..3566
                     /label=6xHis
     misc_feature    complement(3589..3606)
                     /label=bGH-R (genewiz)
     polyA_signal    3595..3819
                     /label=bGH poly(A) signal
     primer_bind     complement(3890..3906)
                     /label=M13 rev
     protein_bind    3914..3930
                     /label=lac operator
     promoter        complement(3938..3968)
                     /label=lac promoter
     misc_feature    3983..4007
                     /label=ACK21
     protein_bind    3983..4004
                     /label=CAP binding site
     rep_origin      complement(4292..4880)
                     /direction=LEFT
                     /label=ori
     misc_feature    4292..4316
                     /label=ACK10
     CDS             complement(5051..5911)
                     /label=AmpR
     CDS             complement(5051..5911)
                     /codon_start=1
                     /label=Translation 5911-5051
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5912..6016)
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
      661 gcaagtaact tcactcaatt tgtgcttgta gacaacggag gtaccggaga tgtaacagta
      721 gcacccagta atttcgcaaa cggcgtggcg gaatggatta gttcaaattc aagatctcag
      781 gcctataagg tcacttgctc cgtcaggcag tctagtgccc aaaagcgcaa atacacgatt
      841 aaagtagagg taccgaaggt cgctacgcaa actgttgggg gagttgaact tccagtggct
      901 gcttggcgct catacctgaa catggaactg actatcccca tattcgctac taactccgat
      961 tgtgaactca tagtaaaagc tatgcaggga ttgctcaagg atggcaaccc cattccaagt
     1021 gccatagcag cgaactccgg gatatattct tctggaggat ctagcggagg atcctctggc
     1081 agcgagacac caggaacaag cgagtcagca acaccagaga gcagtggcgg cagcagcggc
     1141 ggcagcagtt caaagactgg gcctgtcgcc gtcgatccaa ccctgcgccg ccggattgaa
     1201 cctcacgagt ttgaagtgtt ctttgacccc cgggagctga gaaaggagac atgcctgctg
     1261 tacgagatca actggggagg caggcactcc atctggaggc acacctctca gaacacaaat
     1321 aagcacgtgg aggtgaactt catcgagaag tttaccacag agcggtactt ctgccccaat
     1381 accagatgta gcatcacatg gtttctgagc tggtcccctt gcggagagtg tagcagggcc
     1441 atcaccgagt tcctgtccag atatccaaat gtgacactgt ttatctacat cgccaggctg
     1501 tatcacctgg caaacccaag gaataggcag ggcctgcgcg atctgatcag ctccggcgtg
     1561 accatccaga tcatgacaga gcaggagtcc ggctactgct ggcacaactt cgtgaattat
     1621 tctcctagca acgagtccca ctggcctagg tacccacacc tgtgggtgcg cctgtacgtg
     1681 ctggagctgt attgcatcat cctgggcctg cccccttgtc tgaatatcct gcggagaaag
     1741 cagagccagc tgacctcctt tacaatcgcc ctgcagtctt gtcactatca gaggctgcca
     1801 ccccacatcc tgtgggccac aggcctgaag tctggcggat ctagcggagg atcctctggc
     1861 agcgagacac caggaacaag cgagtcagca acaccagaga gcagtggcgg cagcagcggc
     1921 ggcagcacta atctgagcga catcattgag aaggagactg ggaaacagct ggtcattcag
     1981 gagtccatcc tgatgctgcc tgaggaggtg gaggaagtga tcggcaacaa gccagagtct
     2041 gacatcctgg tgcacaccgc ctacgacgag tccacagatg agaatgtgat gctgctgacc
     2101 tctgacgccc ccgagtataa gccttgggcc ctggtcatcc aggattctaa cggcgagaat
     2161 aagatcaaga tgctgagcgg aggatccgga ggatctggag gcagcaccaa cctgtctgac
     2221 atcatcgaga aggagacagg caagcagctg gtcatccagg agagcatcct gatgctgccc
     2281 gaagaagtcg aagaagtgat cggaaacaag cctgagagcg atatcctggt ccataccgcc
     2341 tacgacgaga gtaccgacga aaatgtgatg ctgctgacat ccgacgcccc agagtataag
     2401 ccctgggctc tggtcatcca ggattccaac ggagagaaca aaatcaaaat gctgtctggc
     2461 ggctcaaaaa gaaccgccga cggcagcgaa ttcgagccca agaagaagag gaaagtcgga
     2521 agcggagcta ctaacttcag cctgctgaag caggctggag acgtggagga gaaccctgga
     2581 cctggcagtg gtgagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct
     2641 ggcccaatga aacggacagc cgacggaagc gagttcgagt caccaaagaa gaagcggaaa
     2701 gtcagctctg aggtggagtt ttcccatgag tactggatga gacacgcatt gactctcgca
     2761 aagagggctc gagatgaacg cgaggtgccc gtgggggcag tactcgtgct caacaatcgc
     2821 gtaatcggcg aaggttggaa tagggcaatc ggactccacg accccactgc acatgcggaa
     2881 atcatggccc ttcgacaggg agggcttgtg atgcagaatt atcgacttta tgatgcgacg
     2941 ctgtactcga cgtttgaacc ttgcgtaatg tgcgcgggag ctatgattca ctcccgcatt
     3001 ggacgagttg tattcggtgt tcgcaacgcc aagacgggtg ccgcaggttc actgatggac
     3061 gtgctgcatc atccaggcat gaaccaccgg gtagaaatca cagaaggcat attggcggac
     3121 gaatgtgcgg cgctgttgtg tcgttttttt cgcatgccca ggcgggtctt taacgcccag
     3181 aaaaaagcac aatcctctac tgactctggt ggttcttctg gtggttctag cggcagcgag
     3241 actcccggga cctcagagtc cgccacaccc gaaagttctg gtggttcttc tggtggttct
     3301 aaaagcatca gatgcaagaa ctgcaacaag ctgctgttca aggccgacag cttcgaccac
     3361 atcgagatcc ggtgtcctag atgtaaaaga cacatcatca tgctgaacgc ctgcgagcac
     3421 cccaccgaga agcactgcgg caagcgggaa aagatcaccc acagcgacga gacagtgcgg
     3481 tacggttcaa aaagaactgc cgacggctct gagtttgaac ccaagaagaa aaggaaagtc
     3541 taaccggtca tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc
     3601 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3661 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3721 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3781 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3841 gctggggctc gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3901 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3961 agtgtaaagc ctaggatgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4021 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4081 cgggaagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4141 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4201 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4261 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4321 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4381 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4441 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4501 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4561 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4621 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4681 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4741 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4801 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4861 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacactcagt
     4921 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4981 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5041 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5101 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5161 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5221 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5281 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5341 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5401 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5461 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5521 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5581 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5641 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5701 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5761 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5821 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5881 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5941 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6001 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6061 atcgatctcc cgatccccta gggtcgactc tcagtacaat ctgctctgat gccgcatagt
     6121 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     6181 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     6241 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgtt
//
