LOCUS       Exported                8957 bp ds-DNA     circular SYN 04-JUN-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8957)
  AUTHORS   Institute for Molecular Bioscience
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Nov 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8957
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        6..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     256..275
                     /label=IN_F_PLVIP_seq
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     489..512
                     /label=IN_F1_pLV-LifeAct_seq
     CDS             584..634
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     primer_bind     643..668
                     /label=IN_F_AgeI-TagRFPT
     primer_bind     648..674
                     /label=IN_F_XhoI_TagRFP-T
     CDS             653..1381
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELY"
     primer_bind     complement(1358..1390)
                     /label=IN_R_TagRFPT-BsrGI
     primer_bind     complement(1364..1381)
                     /label=IN_R_TagRFP-T_XbaI
     misc_feature    1475..2027
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     1798..1815
                     /label=IN_F4_pLV-LifeAct_seq
     CDS             2054..2653
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     2375..2392
                     /label=IN_F5_pLV-LifeAct_seq
     LTR             3546..3778
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3850..3971
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3990..4146
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      4012..4032
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(4167..4185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4195..4211)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4353..4808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4834..4938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4939..5799
                     /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"
     rep_origin      5970..6558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6846..6867
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6882..6912
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6920..6936
                     /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)."
     primer_bind     6944..6960
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6981..6999
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     LTR             7254..7434
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7481..7606
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    8099..8332
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    8823..8940
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
       61 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      121 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      181 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      241 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      301 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      361 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      421 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      481 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
      541 cgctaccgga ctcagatctc gagctcaagc ttcgaattcc accatgggcg tggccgacct
      601 gatcaagaag ttcgagagca tcagcaagga ggagggggat ccaccggtcg agatggtgtc
      661 taagggcgaa gagctgatta aggagaacat gcacatgaag ctctacatgg agggcaccgt
      721 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
      781 gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
      841 taccagcttc atgtacggca gcagaacctt catcaaccac acccagggca tccccgactt
      901 ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg
      961 gggcgtgctg accgctaccc aggacaccag cctccaggac ggctgcctca tctacaacgt
     1021 caagatcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg
     1081 ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc ggcctggaag gcagaaccga
     1141 catggccctg aagctcgtgg gcgggggcca cctgatctgc aacttcaaga ccacatacag
     1201 atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactatgtgg accacagact
     1261 ggaaagaatc aaggaggccg acaaagagac ctacgtcgag cagcacgagg tggctgtggc
     1321 cagatactgc gacctcccta gcaaactggg gcacaaactt aatggcatgg acgagctgta
     1381 ctaatgtaca gcatcttgag tcccttttta ccgctgttac caattttctt ttgtctttgg
     1441 gtaaacccgg gaattccgcc cctctccccc cctaacgtta ctggccgaag ccgcttggaa
     1501 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     1561 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     1621 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     1681 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     1741 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     1801 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     1861 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     1921 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     1981 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg atatggccac
     2041 aacggtcgcc accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
     2101 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
     2161 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
     2221 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
     2281 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
     2341 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
     2401 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
     2461 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
     2521 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
     2581 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
     2641 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
     2701 ccatgcatgt attcaatcta agcaggcttt cactttctcg ccaacttaca aggcctttct
     2761 gtgtaaacaa tacctgaacc tttaccccgt tgcccggcaa cggccaggtc tgtgccaagt
     2821 gtttgctgac gcaaccccca ctggctgggg cttggtcatg ggccatcagc gcatgcgtgg
     2881 aaccttttcg gctcctctgc cgatccatac tgcggaactc ctagccgctt gttttgctcg
     2941 cagcaggtct ggagcaaaca ttatcgggac tgataactct gttgtcctat cccgcaaata
     3001 tacatcgttt ccatggctgc taggctgtgc tgccaactgg atcctgcgcg ggacgtcctt
     3061 tgtttacgtc ccgtcggcgc tgaatcctgc ggacgaccct tctcggggtc gcttgggact
     3121 ctctcgtccc cttctccgtc tgccgttccg accgaccacg gggcgcacct ctctttacgc
     3181 ggactccccg tctgtgcctt ctcatctgcc ggaccgtgtg cacttcgctt cacctctgca
     3241 cgtcgcatgg agaccaccgt gaacgcccac caaatattgc ccaaggtctt acataagagg
     3301 actcttggac tctcagcaat gtcaacgacc gaccttgagg catacttcaa agactgtttg
     3361 tttaaagact gggaggagtt gggggaggag attaggttaa aggtctttgt actaggaggc
     3421 tgtaggcata aattggtctg cgcaccagca ccatggcgca atcactagag cggggtacct
     3481 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     3541 ggactggaag ggctaattca ctcccaacga agacaagatc tgctttttgc ttgtactggg
     3601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     3661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     3721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     3781 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     3841 agtgagagga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3901 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3961 aatgtatctt atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc
     4021 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4081 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4141 gaggcctagg gacgtaccca attcgcccta tagtgagtcg tattacgcgc gctcactggc
     4201 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4261 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4321 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4381 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4441 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4501 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4561 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4621 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4681 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4741 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     4801 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4861 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4921 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4981 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5041 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5101 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5161 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5221 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5281 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5341 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5401 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5461 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5521 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5581 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5641 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5701 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5761 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5821 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5881 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6601 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6661 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6721 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     6781 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6841 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     6901 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     6961 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctgcaa
     7021 gcttaatgta gtcttatgca atactcttgt agtcttgcaa catggtaacg atgagttagc
     7081 aacatgcctt acaaggagag aaaaagcacc gtgcatgccg attggtggaa gtaaggtggt
     7141 acgatcgtgc cttattagga aggcaacaga cgggtctgac atggattgga cgaaccactg
     7201 aattgccgca ttgcagagat attgtattta agtgcctagc tcgatacata aacgggtctc
     7261 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
     7321 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
     7381 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     7441 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     7501 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     7561 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     7621 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     7681 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     7741 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     7801 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     7861 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     7921 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
     7981 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
     8041 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
     8101 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
     8161 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
     8221 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
     8281 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
     8341 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
     8401 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
     8461 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
     8521 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
     8581 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
     8641 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
     8701 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
     8761 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct cgacggtatc
     8821 ggttttaaaa gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata
     8881 atagcaacag acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt
     8941 atcgataagc ttgggag
//
