LOCUS       Exported                8296 bp ds-DNA     circular SYN 30-JUL-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8296)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2020 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8296
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
                     /note="color: #ffe4c4"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
                     /note="color: #ffe4c4"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
                     /note="color: #993366; direction: RIGHT"
     regulatory      1418..1427
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             1430..1450
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             1451..3076
                     /codon_start=1
                     /product="green-emitting variant of click beetle 
                     luciferase"
                     /label=CBG99luc
                     /note="human codon-optimized"
                     /note="color: #e1ff2f"
                     /translation="MVKREKNVIYGPEPLHPLEDLTAGEMLFRALRKHSHLPQALVDVV
                     GDESLSYKEFFEATVLLAQSLHNCGYKMNDVVSICAENNTRFFIPVIAAWYIGMIVAPV
                     NESYIPDELCKVMGISKPQIVFTTKNILNKVLEVQSRTNFIKRIIILDTVENIHGCESL
                     PNFISRYSDGNIANFKPLHFDPVEQVAAILCSSGTTGLPKGVMQTHQNICVRLIHALDP
                     RVGTQLIPGVTVLVYLPFFHAFGFSITLGYFMGGLRVIMFRRFDQEAFLKAIQDYEVRS
                     VINVPSVILFLSKSPLVDKYDLSSLRELCCGAAPLAKEVAEVAAKRLNLPGIRCGFGLT
                     ESTSANIHSLGDEFKSGSLGRVTPLMAAKIADRETGKALGPNQVGELCIKGPMVSKGYV
                     NNVEATKEAIDDDGWLHSGDFGYYDEDEHFYVVDRYKELIKYKGSQVAPAELEEILLKN
                     PCIRDVAVVGIPDLEAGELPSAFVVKQPGKEITAKEVYDYLAERVSHTKYLRGGVRFVD
                     SIPRNVTGKITRKELLKQLLEKAGG"
     RBS             2401..2409
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
                     /note="color: #a6acb3"
     CDS             3101..3430
                     /codon_start=1
                     /label=FRA1 PEST domain
                     /note="color: #993366"
                     /translation="LVLEAHRPICKIPEGAKEGDTGSTSGTSSPPAPCRPVPCISLSPG
                     PVLEPEALHTPTLMTTPSLTPFTPSLVFTYPSTPEPCASAHRKSSSSSGDPSSDPLGSP
                     TLLAL"
     promoter        3441..3940
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             3961..4560
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             4660..5174
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
                     /note="color: #ffe4c4"
     primer_bind     complement(5343..5359)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5367..5383
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5391..5421)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5391 .. 5397 / #ffffff / -10
                        2: 5398 .. 5415 / #ffffff
                        3: 5416 .. 5421 / #ffffff / -35"
     protein_bind    5436..5457
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5745..6333)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6504..7364)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6504 .. 7295 / #ccffcc
                        2: 7296 .. 7364 / #ccffcc / signal sequence
                     Cleavage site after base 7295"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7365..7469)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tagatctgcc accatggccc ccaagaagaa
     1441 gaggaaggtg atggtgaagc gtgagaaaaa tgtcatctat ggccctgagc ctctccatcc
     1501 tttggaggat ttgactgccg gcgaaatgct gtttcgtgct ctccgcaagc actctcattt
     1561 gcctcaagcc ttggtcgatg tggtcggcga tgaatctttg agctacaagg agttttttga
     1621 ggcaaccgtc ttgctggctc agtccctcca caattgtggc tacaagatga acgacgtcgt
     1681 tagtatctgt gctgaaaaca atacccgttt cttcattcca gtcatcgccg catggtatat
     1741 cggtatgatc gtggctccag tcaacgagag ctacattccc gacgaactgt gtaaagtcat
     1801 gggtatctct aagccacaga ttgtcttcac cactaagaat attctgaaca aagtcctgga
     1861 agtccaaagc cgcaccaact ttattaagcg tatcatcatc ttggacactg tggagaatat
     1921 tcacggttgc gaatctttgc ctaatttcat ctctcgctat tcagacggca acatcgcaaa
     1981 ctttaaacca ctccacttcg accctgtgga acaagttgca gccattctgt gtagcagcgg
     2041 tactactgga ctcccaaagg gagtcatgca gacccatcaa aacatttgcg tgcgtctgat
     2101 ccatgctctc gatccacgcg tgggcactca gctgattcct ggtgtcaccg tcttggtcta
     2161 cttgcctttc ttccatgctt tcggctttag cattactttg ggttacttta tgggcggtct
     2221 ccgcgtgatt atgttccgcc gttttgatca ggaggctttc ttgaaagcca tccaagatta
     2281 tgaagtccgc agtgtcatca acgtgcctag cgtgatcctg tttttgtcta agagcccact
     2341 cgtggacaag tacgacttgt cttcactgcg tgaattgtgt tgcggtgccg ctccactggc
     2401 taaggaggtc gctgaagtgg ccgccaaacg cttgaatctt ccagggattc gttgtggctt
     2461 cggcctcacc gaatctacca gcgctaacat tcactctctc ggggatgagt ttaagagcgg
     2521 ctctttgggc cgtgtcactc cactcatggc tgctaagatc gctgatcgcg aaactggtaa
     2581 ggctttgggc ccgaaccaag tgggcgagct gtgtatcaaa ggccctatgg tgagcaaggg
     2641 ttatgtcaat aacgttgaag ctaccaagga ggccatcgac gacgacggct ggttgcattc
     2701 tggtgatttt ggatattacg acgaagatga gcatttttac gtcgtggatc gttacaagga
     2761 gctgatcaaa tacaagggta gccaggttgc tccagctgag ttggaggaga ttctgttgaa
     2821 aaatccatgc attcgcgatg tcgctgtggt cggcattcct gatctggagg ccggcgaact
     2881 gccttctgct ttcgttgtca agcagcctgg taaagaaatt accgccaaag aagtgtatga
     2941 ttacctggct gaacgtgtga gccatactaa gtacttgcgt ggcggcgtgc gttttgttga
     3001 ctccatccct cgtaacgtaa caggcaaaat tacccgcaag gagctgttga aacaattgtt
     3061 ggagaaggcc ggcggtttgt acaaaggcag cggcggatcc ctggtgctgg aagcccaccg
     3121 acccatctgc aaaatcccgg aaggagccaa ggagggggac acaggcagta ccagtggcac
     3181 cagcagccca ccagccccct gccgccctgt accttgtatc tccctttccc cagggcctgt
     3241 gcttgaacct gaggcactgc acacccccac actcatgacc acaccctccc taactccttt
     3301 cacccccagc ctggtcttca cctaccccag cactcctgag ccttgtgcct cagctcatcg
     3361 caagagtagc agcagcagcg gagacccatc ctctgacccc cttggctctc caaccctcct
     3421 cgctttgtga gaattctacc gggtagggga ggcgcttttc ccaaggcagt ctggagcatg
     3481 cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc ctcgcacaca
     3541 ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct tcgcgccacc
     3601 ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac
     3661 gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca ccgctgagca
     3721 atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct tcgctttctg
     3781 ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg
     3841 ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc
     3901 tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc caagcttacc
     3961 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     4021 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     4081 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     4141 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     4201 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     4261 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     4321 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     4381 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     4441 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     4501 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     4561 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tgcatcgata
     4621 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     4681 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     4741 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     4801 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     4861 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     4921 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     4981 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     5041 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     5101 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     5161 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     5221 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     5281 ctcctgaaat agttcattat ggacagcgca gaagagctgg ggagaattaa ttcgtaatca
     5341 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5401 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5461 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5521 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5581 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5641 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5701 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5761 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5821 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5881 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5941 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6001 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6061 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6121 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6181 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6241 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6301 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6361 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6421 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6481 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6541 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6601 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6661 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6721 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6781 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6841 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6901 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6961 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7021 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7081 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7141 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7201 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7261 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7321 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7381 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7441 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     7501 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     7561 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     7621 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     7681 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     7741 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     7801 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     7861 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     7921 gggttttccc agtcacgacg ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga
     7981 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     8041 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     8101 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggcgat
     8161 tagtccaatt tgttaaagac aggatatcag tggtccaggc tctagttttg actcaacaat
     8221 atcaccagct gaagcctata gagtacgagc catagataaa ataaaagatt ttatttagtc
     8281 tccagaaaaa gggggg
//
