LOCUS       Exported                7853 bp DNA     circular SYN 11-JAN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7853)
  AUTHORS   nolanmaier
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 2021 from SnapGene Viewer 5.3.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7853
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     primer_bind     925..948
                     /label=oNM081
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated MMLV gag gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBabe-5'-primer
     CDS             1360..1737
                     /codon_start=1
                     /label=hH2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     primer_bind     1768..1798
                     /label=oNM1248
     CDS             1780..2472
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     1780..1795
                     /label=cterm_mC_1F
     primer_bind     complement(2452..2473)
                     /label=mCherry gibson REV
     primer_bind     complement(2453..2472)
                     /label=gfp_2R
     primer_bind     complement(2461..2505)
                     /label=oNM1249
     primer_bind     2499..2519
                     /label=oNM162
     CDS             2506..3501
                     /codon_start=1
                     /label=hMeikin (1-332)
                     /translation="MWPLRVYTRKKREGQRLNLTPTPDLGSPAKAEAPPGSKRKGKVHG
                     LSKIAEKAERSRQGGSGSGPFSPRLGVTGEKSLQENRSSEDTQDEKIASLRESVTDDLQ
                     VDSSSSNSELVSGLSLHHGMASSLLSYSVTDSYAEYKSFEESFPSPELFRKSDYLDWEC
                     PNLEEHMQWKNSTLLDTSKAVAIEKAPQFSNVSAIFSTSSEDYQKCHRKTVMTVADQNV
                     SPKAKCASNSESDNAACEILLAEKTCPSTPEKTKKKKTNSSTPGKKNRGLLTSTPSSET
                     AGFVIDLSSVQKASFEELFPNVSNYVNSNEIVPVSSLQENSSNEFPANASE"
     primer_bind     complement(3480..3533)
                     /label=oNM1175
     primer_bind     3522..3565
                     /label=cterm_mC_1F
     CDS             3550..4254
                     /codon_start=2
                     /label=NeoGreen Human optimized
                     /note="NeoGreen Human optimized"
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      3802..3811
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     3909..3919
                     /label=cterm_mC_1F
     primer_bind     complement(4234..4255)
                     /label=mCherry gibson REV
     primer_bind     complement(4235..4254)
                     /label=gfp_2R
     primer_bind     complement(4243..4255)
                     /label=oNM1249
     primer_bind     complement(4269..4289)
                     /label=pBabe-3'-primer
     promoter        4274..4603
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4454..4589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4516..4535
                     /label=SV40promoter-primer-Forward
     promoter        4618..4665
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4684..5115
                     /codon_start=1
                     /label=BlastR
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVRTVMDSRRQLGFVNCCPLVMCGRAKHFIDKIKDFI"
     LTR             5134..5727
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(6099..6687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6865..7725)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7726..7830)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggcgccacca tgccagagcc agcgaagtct
     1381 gctcccgccc cgaaaaaggg ctccaagaag gcggtgacta aggcgcagaa gaaaggcggc
     1441 aagaagcgca agcgcagccg caaggagagc tattccatct atgtgtacaa ggttctgaag
     1501 caggtccacc ctgacaccgg catttcgtcc aaggccatgg gcatcatgaa ttcgtttgtg
     1561 aacgacattt tcgagcgcat cgcaggtgag gcttcccgcc tggcgcatta caacaagcgc
     1621 tcgaccatca cctccaggga gatccagacg gccgtgcgcc tgctgctgcc tggggagttg
     1681 gccaagcacg ccgtgtccga gggtactaag gccatcacca agtacaccag cgctaagggt
     1741 ggtggcggtt caggaagcgg gtccggctct gggcgcgccg tgagcaaggg cgaggcagtg
     1801 atcaaggagt tcatgcggtt caaggtgcac atggagggct ccatgaacgg ccacgagttc
     1861 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag
     1921 gtgaccaagg gtggccccct gcccttctcc tgggacatcc tgtcccctca gttcatgtac
     1981 ggctccaggg ccttcaccaa gcaccccgcc gacatccccg actactataa gcagtccttc
     2041 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg
     2101 acccaggaca cctccctgga ggacggcacc ctgatctaca aggtgaagct ccgcggcacc
     2161 aacttccctc ctgacggccc cgtaatgcag aagaagacaa tgggctggga agcgtccacc
     2221 gagcggttgt accccgagga cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg
     2281 aaggacggcg gccgctacct ggcggacttc aagaccacct acaaggccaa gaagcccgtg
     2341 cagatgcccg gcgcctacaa cgtcgaccgc aagttggaca tcacctccca caacgaggac
     2401 tacaccgtgg tggaacagta cgaacgctcc gagggccgac actccaccgg cggcatggac
     2461 gagctgtaca agtccggagg gggctctggt ggaggcagcc tcgagatgtg gcccctgagg
     2521 gtgtacacca gaaagaagag ggagggccag agactgaacc tgacacccac acctgacctg
     2581 ggctcccctg ccaaggctga agctcctccc ggcagcaaga ggaaaggcaa ggtgcacggc
     2641 ctgagcaaga tcgctgagaa agccgagaga agcaggcagg gcggatccgg aagcggacct
     2701 ttcagcccca ggctcggagt gaccggcgag aagtccctgc aggagaacag gtccagcgag
     2761 gacacccagg acgagaagat tgcttccctg agggagagcg tgaccgacga cctgcaggtg
     2821 gactccagca gctccaactc cgaactggtg tccggactga gcctgcatca cggcatggcc
     2881 tcctccctgc tgtcctatag cgtgaccgat tcctacgccg agtacaagag cttcgaggag
     2941 tccttcccct cccctgagct gttcaggaag tccgactacc tggactggga gtgccccaat
     3001 ctggaggagc acatgcagtg gaagaacagc accctgctcg acacctccaa ggccgtggct
     3061 atcgagaagg ctccccagtt cagcaacgtc agcgccatct tctccaccag cagcgaggac
     3121 taccagaagt gccacaggaa gaccgtgatg accgtggccg atcagaacgt gtcccccaag
     3181 gccaagtgcg ccagcaatag cgagagcgac aacgctgcct gtgaaatcct gctggccgag
     3241 aagacatgcc ccagcacccc cgagaagacc aagaagaaga agaccaacag cagcacacct
     3301 ggcaagaaga acagaggact gctgacctcc acacctagct ccgagaccgc cggctttgtg
     3361 atcgacctga gctccgtgca gaaggcctcc tttgaggagc tgttccccaa cgtgagcaac
     3421 tacgtgaaca gcaacgagat cgtccccgtg tccagcctcc aggagaacag ctccaacgag
     3481 ttccccgcca acgcctccga gggaagcggc tccgggagtg gatccggacc ggtaggcggc
     3541 ggtgggtccg tgagcaaggg cgaggaggac aacatggcca gcctgcccgc cacccacgag
     3601 ctgcacatct tcggcagcat caacggcgtg gacttcgaca tggtgggcca gggcaccggc
     3661 aaccccaacg acggctacga ggagctgaac ctgaagagca ccaagggcga cctgcagttc
     3721 agcccctgga ttctggtgcc ccacatcggc tacggcttcc accagtacct gccctacccc
     3781 gacggcatga gccccttcca ggccgccatg gtggacggca gcggctacca ggtgcaccgc
     3841 accatgcagt tcgaggacgg cgccagcctg accgtgaact accgctacac ctacgagggc
     3901 agccacatca agggcgaggc ccaggtgaag ggcaccggct tccccgccga cggccccgtg
     3961 atgaccaaca gcctgaccgc cgccgactgg tgccgcagca agaagaccta ccccaacgac
     4021 aagaccatca tcagcacctt caagtggagc tacaccaccg gcaacggcaa gcgctaccgc
     4081 agcaccgccc gcaccaccta caccttcgcc aagcccatgg ccgccaacta cctgaagaac
     4141 cagcccatgt acgtgttccg caagaccgag ctgaagcaca gcaagaccga gctgaacttc
     4201 aaggagtggc agaaggcctt caccgacgtg atgggcatgg acgagctgta caagtaagtc
     4261 gaccctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     4321 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4381 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4441 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4501 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4561 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagcttg agcacgtgtt
     4621 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     4681 accatggcca agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca
     4741 atcaacagca tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac
     4801 ggccgcatct tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc
     4861 gtggtgctgg gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc
     4921 ggaaatgaga acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat
     4981 ctgcatcctg ggatcaaagc catagtgagg acagtgatgg acagccgacg gcagttggga
     5041 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta agcacttcat cgataaaata
     5101 aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg
     5161 caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga
     5221 gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc
     5281 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg
     5341 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     5401 tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg
     5461 ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg
     5521 cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg
     5581 gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct
     5641 tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg
     5701 actacccgtc agcgggggtc tttcatttcc gacttgtggt ctcgctgcct tgggagggtc
     5761 tcctctgagt gattgactac ccgtcagcgg gggtcttcac atgcagcatg tatcaaaatt
     5821 aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg
     5881 ccaacgcgcg gggagaggcg gtttgcgtat tggcgctctt ccgcttcctc gctcactgac
     5941 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6001 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6061 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     6541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6661 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6721 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6781 ttcacctaga tccttttaaa ttaaaaatga agtttgcggc cgcaaatcaa tctaaagtat
     6841 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     6901 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     6961 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7021 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7081 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7141 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7201 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7261 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7321 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7381 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7441 atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata ataccgcgcc
     7501 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7561 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7621 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     7681 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     7741 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     7801 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
