LOCUS       Exported                8387 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 8387)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2020 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8387
                     /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"
     CDS             1436..1456
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /note="color: #993366"
                     /translation="PKKKRKV"
     CDS             1457..3082
                     /codon_start=1
                     /product="red-emitting variant of click beetle luciferase"
                     /label=CBRluc
                     /note="human codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVKREKNVIYGPEPLHPLEDLTAGEMLFRALRKHSHLPQALVDVV
                     GDESLSYKEFFEATVLLAQSLHNCGYKMNDVVSICAENNTRFFIPVIAAWYIGMIVAPV
                     NESYIPDELCKVMGISKPQIVFTTKNILNKVLEVQSRTNFIKRIIILDTVENIHGCESL
                     PNFISRYSDGNIANFKPLHFDPVEQVAAILCSSGTTGLPKGVMQTHQNICVRLIHALDP
                     RYGTQLIPGVTVLVYLPFFHAFGFHITLGYFMVGLRVIMFRRFDQEAFLKAIQDYEVRS
                     VINVPSVILFLSKSPLVDKYDLSSLRELCCGAAPLAKEVAEVAAKRLNLPGIRCGFGLT
                     ESTSAIIQTLGDEFKSGSLGRVTPLMAAKIADRETGKALGPNQVGELCIKGPMVSKGYV
                     NNVEATKEAIDDDGWLHSGDFGYYDEDEHFYVVDRYKELIKYKGSQVAPAELEEILLKN
                     PCIRDVAVVGIPDLEAGELPSAFVVKQPGTEITAKEVYDYLAERVSHTKYLRGGVRFVD
                     SIPRNVTGKITRKELLKQLLVKAGG"
     RBS             2407..2415
                     /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             3092..3220
                     /codon_start=1
                     /label=d2 PEST domain
                     /note="color: #993366"
                     /translation="LESHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV"
     promoter        3255..3584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3435..3570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             3658..4683
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /note="color: #ccffcc"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     LTR             4751..5265
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
                     /note="color: #ffe4c4"
     primer_bind     complement(5434..5450)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5458..5474
                     /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(5482..5512)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5482 .. 5488 / #ffffff / -10
                        2: 5489 .. 5506 / #ffffff
                        3: 5507 .. 5512 / #ffffff / -35"
     protein_bind    5527..5548
                     /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(5836..6424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6595..7455)
                     /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: 6595 .. 7386 / #ccffcc
                        2: 7387 .. 7455 / #ccffcc / signal sequence
                     Cleavage site after base 7386"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7456..7560)
                     /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 tagatctcct aggggatcca tggcccccaa
     1441 gaagaagagg aaggtgatgg taaagcgtga gaaaaatgtc atctatggcc ctgagcctct
     1501 ccatcctttg gaggatttga ctgccggcga aatgctgttt cgtgctctcc gcaagcactc
     1561 tcatttgcct caagccttgg tcgatgtggt cggcgatgaa tctttgagct acaaggagtt
     1621 ttttgaggca accgtcttgc tggctcagtc cctccacaat tgtggctaca agatgaacga
     1681 cgtcgttagt atctgtgctg aaaacaatac ccgtttcttc attccagtca tcgccgcatg
     1741 gtatatcggt atgatcgtgg ctccagtcaa cgagagctac attcccgacg aactgtgtaa
     1801 agtcatgggt atctctaagc cacagattgt cttcaccact aagaatattc tgaacaaagt
     1861 cctggaagtc caaagccgca ccaactttat taagcgtatc atcatcttgg acactgtgga
     1921 gaatattcac ggttgcgaat ctttgcctaa tttcatctct cgctattcag acggcaacat
     1981 cgcaaacttt aaaccactcc acttcgaccc tgtggaacaa gttgcagcca ttctgtgtag
     2041 cagcggtact actggactcc caaagggagt catgcagacc catcaaaaca tttgcgtgcg
     2101 tctgatccat gctctcgatc cacgctacgg cactcagctg attcctggtg tcaccgtctt
     2161 ggtctacttg cctttcttcc atgctttcgg ctttcatatt actttgggtt actttatggt
     2221 cggtctccgc gtgattatgt tccgccgttt tgatcaggag gctttcttga aagccatcca
     2281 agattatgaa gtccgcagtg tcatcaacgt gcctagcgtg atcctgtttt tgtctaagag
     2341 cccactcgtg gacaagtacg acttgtcttc actgcgtgaa ttgtgttgcg gtgccgctcc
     2401 actggctaag gaggtcgctg aagtggccgc caaacgcttg aatcttccag ggattcgttg
     2461 tggcttcggc ctcaccgaat ctaccagtgc gattatccag actctcgggg atgagtttaa
     2521 gagcggctct ttgggccgtg tcactccact catggctgct aagatcgctg atcgcgaaac
     2581 tggtaaggct ttgggcccga accaagtggg cgagctgtgt atcaaaggcc ctatggtgag
     2641 caagggttat gtcaataacg ttgaagctac caaggaggcc atcgacgacg acggctggtt
     2701 gcattctggt gattttggat attacgacga agatgagcat ttttacgtcg tggatcgtta
     2761 caaggagctg atcaaataca agggtagcca ggttgctcca gctgagttgg aggagattct
     2821 gttgaaaaat ccatgcattc gcgatgtcgc tgtggtcggc attcctgatc tggaggccgg
     2881 cgaactgcct tctgctttcg ttgtcaagca gcctggtaca gaaattaccg ccaaagaagt
     2941 gtatgattac ctggctgaac gtgtgagcca tactaagtac ttgcgtggcg gcgtgcgttt
     3001 tgttgactcc atccctcgta acgtaacagg caaaattacc cgcaaggagc tgttgaaaca
     3061 attgttggtg aaggccggcg gtttgtacaa actcgagagc catggcttcc cgccggaggt
     3121 ggaggagcag gatgatggca cgctgcccat gtcttgtgcc caggagagcg ggatggaccg
     3181 tcaccctgca gcctgtgctt ctgctaggat caatgtgtag gaattcgcca gcacagtggt
     3241 cgaccctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3301 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3421 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3481 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3541 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctt gccagctggg
     3601 gcgccctctg gtaaggttgg gaagccctgc agatctgatc ccggggggca atgagatatg
     3661 aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc tgatcgaaaa gttcgacagc
     3721 gtctccgacc tgatgcagct ctcggagggc gaagaatctc gtgctttcag cttcgatgta
     3781 ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg atggtttcta caaagatcgt
     3841 tatgtttatc ggcactttgc atcggccgcg ctcccgattc cggaagtgct tgacattggg
     3901 gaatttagcg agagcctgac ctattgcatc tcccgccgtg cacagggtgt cacgttgcaa
     3961 gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg tcgcggaggc catggatgcg
     4021 atcgctgcgg ccgatcttag ccagacgagc gggttcggcc cattcggacc gcaaggaatc
     4081 ggtcaataca ctacatggcg tgatttcata tgcgcgattg ctgatcccca tgtgtatcac
     4141 tggcaaactg tgatggacga caccgtcagt gcgtccgtcg cgcaggctct cgatgagctg
     4201 atgctttggg ccgaggactg ccccgaagtc cggcacctcg tgcacgcgga tttcggctcc
     4261 aacaatgtcc tgacggacaa tggccgcata acagcggtca ttgactggag cgaggcgatg
     4321 ttcggggatt cccaatacga ggtcgccaac atcttcttct ggaggccgtg gttggcttgt
     4381 atggagcagc agacgcgcta cttcgagcgg aggcatccgg agcttgcagg atcgccgcgg
     4441 ctccgggcgt atatgctccg cattggtctt gaccaactct atcagagctt ggttgacggc
     4501 aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg caatcgtccg atccggagcc
     4561 gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg ccgtctggac cgatggctgt
     4621 gtagaagtac tcgccgatag tggaaaccga cgccccagca ctcgtccgag ggcaaaggaa
     4681 tagagtagat gccgaccggg atctatcgat aaaataaaag attttattta gtctccagaa
     4741 aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt
     4801 ttgcaaggca tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca
     4861 gagagacagc agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
     4921 cagggccaag aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc
     4981 atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa
     5041 ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga
     5101 gcccacaacc cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg
     5161 tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag
     5221 ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcatgggta acagtttctt
     5281 gaagttggag aacaacattc tgagggtagg agtcgaatat taagtaatcc tgactcaatt
     5341 agccactgtt ttgaatccac atactccaat actcctgaaa tagttcatta tggacagcgc
     5401 agaagagctg gggagaatta attcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5461 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5521 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5581 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5641 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5701 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5761 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5821 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5881 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5941 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6001 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6061 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6121 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6181 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6241 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     6301 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6361 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6421 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6481 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6541 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6601 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6661 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6721 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6781 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6841 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6901 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6961 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7021 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7081 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7141 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7201 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7261 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7321 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7381 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7441 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7501 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7561 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     7621 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     7681 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     7741 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     7801 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     7861 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     7921 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     7981 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     8041 gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     8101 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     8161 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     8221 ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca
     8281 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag
     8341 ccatagataa aataaaagat tttatttagt ctccagaaaa agggggg
//
