LOCUS       Exported                7772 bp ds-DNA     circular SYN 26-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7772)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2024 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7772
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1718..1753
                     /label=CAG-ChRm_F
     primer_bind     1718..1740
                     /label=ChRmine_F
     CDS             1738..2664
                     /codon_start=1
                     /label=ChRmine
                     /translation="MAHAPGTDQMFYVGTMDGWYLDTKLNSVAIGAHWSCFIVLTITTF
                     YLGYESWTSRGPSKRTSFYAGYQEEQNLALFVNFFAMLSYFGKIVADTLGHNFGDVGPF
                     IIGFGNYRYADYMLTCPMLVYDLLYQLRAPYRVSCSAIIFAILMSGVLAEFYAEGDPRL
                     RNGAYAWYGFGCFWFIFAYSIVMSIVAKQYSRLAQLAQDTGAEHSLHVLKFAVFTFSML
                     WILFPLVWAICPRGFGWIDDNWTEVAHCVCDIVAKSCYGFALARFRKTYDEELFRLLEQ
                     LGHDEDEFQKLELDMRLSSNGERLRRLS"
     misc_feature    1819..1896
                     /label=TM1
     misc_feature    1939..1995
                     /label=TM2
     primer_bind     complement(2046..2100)
                     /label=T119V_R
     primer_bind     complement(2046..2098)
                     /label=T119G_R
     primer_bind     complement(2046..2097)
                     /label=T119S_R
     primer_bind     complement(2057..2090)
                     /label=D115M_R1
     primer_bind     complement(2061..2090)
                     /label=D115E_R1
     misc_feature    2071..2124
                     /label=TM3
     primer_bind     2074..2104
                     /label=D115M_F2
     primer_bind     2074..2097
                     /label=D115E_F2
     primer_bind     2086..2135
                     /label=T119V_F
     primer_bind     2087..2132
                     /label=T119G_F
     primer_bind     2087..2132
                     /label=T119S_F
     misc_feature    2143..2196
                     /label=TM4
     primer_bind     complement(2219..2267)
                     /label=G174S_R
     misc_feature    2233..2343
                     /label=TM5
     primer_bind     2241..2298
                     /label=G174S_F
     misc_feature    2359..2433
                     /label=TM6
     misc_feature    2461..2544
                     /label=TM7
     primer_bind     complement(2461..2501)
                     /label=D253E_R
     primer_bind     complement(2469..2517)
                     /label=A256V_R
     primer_bind     complement(2476..2517)
                     /label=S258N_R
     primer_bind     complement(2479..2523)
                     /label=S258E_R
     primer_bind     2488..2529
                     /label=D253E_F
     primer_bind     2495..2538
                     /label=A265V_F
     primer_bind     2501..2544
                     /label=S258E_F
     primer_bind     2501..2544
                     /label=S258N_F
     primer_bind     complement(2659..2672)
                     /label=CAG-ChRm_R
     misc_feature    2674..2733
                     /label=Surface expression
     primer_bind     complement(2712..2734)
                     /label=Fragment.REV
     primer_bind     complement(2730..2749)
                     /label=ChRmine_R
     misc_feature    2736..2742
                     /label=Linker
     CDS             2743..3453
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     CDS             3463..3657
                     /codon_start=1
                     /label=Soma-targeting motif
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     intron          3695..3828
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     misc_feature    3837..4425
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     misc_feature    3863..3882
                     /label=seq primer 5
     CDS             complement(4308..4319)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4458..4569
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     polyA_site      4861..4916
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(5275..5291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5299..5315
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5323..5353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5368..5389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5448..5643
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5494..5629
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5649..5783
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6022..6610)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6781..7641)
                     /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"
     misc_feature    6822..6841
                     /label=Seq primer 8
     misc_feature    7469..7488
                     /label=Seq primer 7
     promoter        complement(7642..7746)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg ggcgagggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcgg taccaccatg
     1741 gcacacgcac caggcaccga ccagatgttc tacgtgggca caatggacgg ctggtatctg
     1801 gataccaagc tgaactccgt ggccatcggc gcccactggt cttgctttat cgtgctgaca
     1861 atcaccacat tctacctggg ctatgagtcc tggacatctc ggggccctag caagagaacc
     1921 tccttttacg ccggctatca ggaggagcag aacctggccc tgtttgtgaa tttctttgcc
     1981 atgctgagct acttcggcaa gatcgtggcc gacacactgg gccacaactt cggcgatgtg
     2041 ggccccttca tcatcggctt cggcaattac aggtatgccg actacatgct gacctgccca
     2101 atgctggtgt acgatctgct gtatcagctg agggccccct atcgcgtgtc ttgtagcgcc
     2161 atcatctttg ccatcctgat gtctggcgtg ctggccgagt tctacgcaga gggcgaccct
     2221 aggctgagga atggcgccta cgcctggtat ggctttggct gtttctggtt tatcttcgcc
     2281 tactctatcg tgatgagcat cgtggccaag cagtatagcc ggctggccca gctggcccag
     2341 gatacaggcg ccgagcacag cctgcacgtg ctgaagttcg ccgtgtttac cttctccatg
     2401 ctgtggattc tgtttcccct ggtgtgggcc atctgcccta gaggcttcgg ctggatcgac
     2461 gataactgga cagaggtggc ccactgcgtg tgcgacatcg tggccaagtc ctgttacggc
     2521 tttgccctgg cccggttcag aaagacctat gatgaggagc tgtttcggct gctggagcag
     2581 ctgggacacg acgaggatga gttccagaag ctggagctgg atatgaggct gagcagcaat
     2641 ggcgagcgcc tgcggagact gtctgcggcc gccaagagca ggatcaccag cgagggcgag
     2701 tacatccccc tggaccagat cgacatcaac gtggtaccgg taatggtgtc taagggcgaa
     2761 gagctgatca aggaaaatat gcgtatgaag gtggtcatgg aaggttcggt caacggccac
     2821 caattcaaat gcacaggtga aggagaaggc aatccgtaca tgggaactca aaccatgagg
     2881 atcaaagtca tcgagggagg acccctgcca tttgcctttg acattcttgc cacgtcgttc
     2941 atgtatggca gccgtacttt tatcaagtac ccgaaaggca ttcctgattt ctttaaacag
     3001 tcctttcctg agggttttac ttgggaaaga gttacgagat acgaagatgg tggagtcgtc
     3061 accgtcatgc aggacaccag ccttgaggat ggctgtctcg tttaccacgt ccaagtcaga
     3121 ggggtaaact ttccctccaa tggtcccgtg atgcagaaga agaccaaggg ttgggagcct
     3181 aatacagaga tgatgtatcc agcagatggt ggtctgaggg gatacactca tatggcactg
     3241 aaagttgatg gtggtggcca tctgtcttgc tctttcgtaa caacttacag gtcaaaaaag
     3301 accgtcggga acatcaagat gcccggtatc catgccgttg atcaccgcct ggaaaggtta
     3361 gaggaaagtg acaatgaaat gttcgtagta caacgcgaac acgcagttgc caagttcgcc
     3421 gggcttggtg gtgggatgga cgagctgtac aaggccctag agcagtccca gcccatcctc
     3481 aacaccaagg agatggcccc gcagagcaag cctccagagg agctggagat gagcagcatg
     3541 cccagccccg tggcccctct gcccgcacgc acggagggcg tcatcgacat gcggagcatg
     3601 tccagcattg acagcttcat cagctgtgcc acggacttcc ctgaagccac cagattcttc
     3661 tgctacgaga acgaggtgta actcgaggcc gcaggtaagt atcaaggtta caagacaggt
     3721 ttaaggagac cacatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat
     3781 aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt gtcgacaatc
     3841 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     3901 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     3961 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     4021 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     4081 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     4141 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     4201 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     4261 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     4321 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     4381 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attcgagctc
     4441 ggtacgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     4501 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     4561 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     4621 cagcaagggg gaggattggg aagacaatag caggccagct tttgttccct ttagggttaa
     4681 ttgcgcgctt ggcgctaatc atggtcatag ctgtttccac tcctcaggtg caggctgcct
     4741 atcagaaggt ggtggctggt gtggccaatg ccctggctca caaataccac tgagatcttt
     4801 ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct
     4861 aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg
     4921 gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt
     4981 tggcaacata tgccatatgc tggctgccat gaacaaaggt ggctataaag aggtcatcag
     5041 tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt
     5101 tagatttttt tttatatttt gtttgtgtta tttttttctt taacatccct aaaattttcc
     5161 ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc
     5221 cctcttctct tatgaagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata
     5281 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     5341 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5401 ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag
     5461 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     5521 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     5581 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     5641 aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     5701 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5761 atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg
     5821 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5881 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5941 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6001 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6061 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6121 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6181 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     6241 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6301 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6361 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6421 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6481 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6541 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6601 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6661 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6721 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6781 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6841 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6901 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6961 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7021 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7081 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7141 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7201 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7261 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7321 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7381 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7441 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7501 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7561 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7621 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7681 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7741 tccgcgcaca tttccccgaa aagtgccacc tg
//
