LOCUS       Exported                7539 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    LV-RFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7539)
  AUTHORS   Beronja S, Livshits G, Williams S, Fuchs E
  TITLE     Rapid functional dissection of genetic networks via tissue-specific 
            transduction and RNAi in mouse embryos.
  JOURNAL   Nat Med. 2010 Jul . 16(7):821-7.
  PUBMED    20526348
REFERENCE   2  (bases 1 to 7539)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7539
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     426..444
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             519..896
                     /codon_start=1
                     /gene="HIST1H2BJ"
                     /product="human histone H2B"
                     /label=H2B
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKV
                     LKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLP
                     GELAKHAVSEGTKAITKYTSAK"
     regulatory      909..918
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             915..1592
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(1045..1063)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1313..1332
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1507..1530
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     LTR             1712..1945
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2017..2138
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2054..2073)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2108..2127
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2178..2313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2240..2259
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2333..2352)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2334..2352)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2362..2379)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2362..2378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2371..2393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2520..2975
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2607..2626)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2817..2838
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3001..3105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3106..3966
                     /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"
     primer_bind     complement(3324..3343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4137..4725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4626..4645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4879..4896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5013..5034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5049..5079
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5087..5103
                     /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     5092..5114
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5111..5127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5111..5127
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     5146..5166
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        5148..5166
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        5194..5420
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             5421..5601
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    5648..5773
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    6266..6499
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             6684..6728
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        7026..7266
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     7026..7046
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     7197..7216
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    7370..7487
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        7536..507
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 ttggggttgc gccttttcca aggcagccct gggtttgcgc agggacgcgg ctgctctggg
       61 cgtggttccg ggaaacgcag cggcgccgac cctgggtctc gcacattctt cacgtccgtt
      121 cgcagcgtca cccggatctt cgccgctacc cttgtgggcc ccccggcgac gcttcctgct
      181 ccgcccctaa gtcgggaagg ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa
      241 gccgcacgtc tcactagtac cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc
      301 cgaccgcgat gggctgtggc caatagcggc tgctcagcag ggcgcgccga gagcagcggc
      361 cgggaagggg cggtgcggga ggcggggtgt ggggcggtag tgtgggccct gttcctgccc
      421 gcgcggtgtt ccgcattctg caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt
      481 gaccgaatca ccgacctctc tccccagggg gatctaccat gccagagcca gcgaagtctg
      541 ctcccgcccc gaaaaagggc tccaagaagg cggtgactaa ggcgcagaag aaaggcggca
      601 agaagcgcaa gcgcagccgc aaggagagct attccatcta tgtgtacaag gttctgaagc
      661 aggtccaccc tgacaccggc atttcgtcca aggccatggg catcatgaac tcgtttgtga
      721 acgacatttt cgagcgcatc gcaggtgagg cttcccgcct ggcgcattac aacaagcgct
      781 cgaccatcac ctccagggag atccagacgg ccgtgcgcct gctgctgcct ggggagttgg
      841 ccaagcacgc cgtgtccgag ggtactaagg ccatcaccaa gtacaccagc gctaaggatc
      901 caccggtcgc caccatggcc tcctccgagg acgtcatcaa ggagttcatg cgcttcaagg
      961 tgcgcatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     1021 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc cccctgccct
     1081 tcgcctggga catcctgtcc cctcagttcc agtacggctc caaggcctac gtgaagcacc
     1141 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     1201 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     1261 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     1321 tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg gatgtacccc gaggacggcg
     1381 ccctgaaggg cgagatcaag atgaggctga agctgaagga cggcggccac tacgacgccg
     1441 aggtcaagac cacctacatg gccaagaagc ccgtgcagct gcccggcgcc tacaagaccg
     1501 acatcaagct ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgagc
     1561 gcgccgaggg ccgccactcc accggcgcct aactcgagac tagtcgacgt gaggatcctc
     1621 tagagtccct aggatgcatc ggtaccttta agaccaatga cttacaaggc agctgtagat
     1681 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     1741 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     1801 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     1861 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     1921 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     1981 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     2041 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     2101 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     2161 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     2221 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     2281 tccagaagta gtgaggaggc ttttttggag gcctagggac gtacccaatt cgccctatag
     2341 tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc
     2401 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
     2461 cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga
     2521 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2581 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2641 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2701 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2761 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2821 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2881 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2941 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg
     3001 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3061 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     3241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     3301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     3361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     3421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     3481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     3541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     3601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     3661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     3721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     3781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     3841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     3901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     3961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     4021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     4081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     4921 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     4981 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     5041 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     5101 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     5161 taaagggaac aaaagctgga gctgcaagct taatgtagtc ttatgcaata ctcttgtagt
     5221 cttgcaacat ggtaacgatg agttagcaac atgccttaca aggagagaaa aagcaccgtg
     5281 catgccgatt ggtggaagta aggtggtacg atcgtgcctt attaggaagg caacagacgg
     5341 gtctgacatg gattggacga accactgaat tgccgcattg cagagatatt gtatttaagt
     5401 gcctagctcg atacataaac gggtctctct ggttagacca gatctgagcc tgggagctct
     5461 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5521 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5581 cagtgtggaa aatctctagc agtggcgccc gaacagggac ttgaaagcga aagggaaacc
     5641 agaggagctc tctcgacgca ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg
     5701 gcggcgactg gtgagtacgc caaaaatttt gactagcgga ggctagaagg agagagatgg
     5761 gtgcgagagc gtcagtatta agcgggggag aattagatcg cgatgggaaa aaattcggtt
     5821 aaggccaggg ggaaagaaaa aatataaatt aaaacatata gtatgggcaa gcagggagct
     5881 agaacgattc gcagttaatc ctggcctgtt agaaacatca gaaggctgta gacaaatact
     5941 gggacagcta caaccatccc ttcagacagg atcagaagaa cttagatcat tatataatac
     6001 agtagcaacc ctctattgtg tgcatcaaag gatagagata aaagacacca aggaagcttt
     6061 agacaagata gaggaagagc aaaacaaaag taagaccacc gcacagcaag cggccgctga
     6121 tcttcagacc tggaggagga gatatgaggg acaattggag aagtgaatta tataaatata
     6181 aagtagtaaa aattgaacca ttaggagtag cacccaccaa ggcaaagaga agagtggtgc
     6241 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
     6301 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
     6361 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
     6421 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
     6481 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
     6541 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
     6601 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
     6661 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
     6721 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
     6781 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
     6841 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
     6901 ccgacaggcc cgaaggaata gaagaagaag gtggagagag agacagagac agatccattc
     6961 gattagtgaa cggatctcga cggtatcgat cacgagacta gcctcgagcg gccgccccct
     7021 tcaccgaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
     7081 gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
     7141 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
     7201 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
     7261 aaggacgaaa caccggcaac aagatgaaga gcaccaactc gagttggtgc tcttcatctt
     7321 gttgttttta attctcgacc tcgagacaaa tggcagtatt catccacaat tttaaaagaa
     7381 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
     7441 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
     7501 gggacagcag agatccactt tggccgcggc tcgaggggg
//
