LOCUS       pLMN-ZsGreen-Neo        8065 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Retroviral vector with a ZsGreen marker, for expressing a short 
            hairpin RNA (shRNA) with an """"UltramiR"""" microRNA scaffold.
ACCESSION   .
VERSION     .
KEYWORDS    pLMN-ZsGreen-Neomycin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8065)
  AUTHORS   transOMIC
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8065)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..515
                     /label=MSCV
                     /note="Murine embryonic stem cell virus promoter including
                     the enhancer and promoter region of PCMV virus and 5' 
                     untranslated region of dl-587rev retrovirus"
     misc_feature    579..920
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             987..1403
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     ncRNA           1419..1540
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     misc_RNA        1541..1609
                     /label=shRNA
                     /note="shRNA"
                     /note="short hairpin RNA"
     gap             1544..1565
                     /estimated_length=22
     gap             1585..1606
                     /estimated_length=22
     ncRNA           1610..1736
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     promoter        1757..2256
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             2270..3070
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    3083..3658
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3659..4351
                     /label=ZsGreen1
                     /note="Zoanthus green fluorescent protein"
     LTR             4458..4972
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem
                     cell virus"
     protein_bind    complement(5134..5150)
                     /label=lac operator
                     /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)."
     promoter        complement(5158..5188)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5203..5224)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5512..6100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6274..7131)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7132..7236)
                     /label=AmpR promoter
     LTR             8064..8065
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc
      301 gcttctcgct tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc
      361 ctcactcggc gcgccagtcc tccgatagac tgcgtcgccc gggtacccgt attcccaata
      421 aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag ggtctcctca
      481 gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga gatttggaga
      541 cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc ggtcgtttcg
      601 tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc ctgcgtctgt
      661 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttctgaa
      721 cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt tgtggcccga
      781 cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt ctggtaggag
      841 acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tggaaccgaa
      901 gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc tgtctgactg
      961 tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta agtttgacct
     1021 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
     1081 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
     1141 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
     1201 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
     1261 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
     1321 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
     1381 ccctcactcc ttctctaggc gccggaatta gatcttgttt gaatgaggct tcagtacttt
     1441 acagaatcgt tgcctgcaca tcttggaaac acttgctggg attacttctt caggttaacc
     1501 caacagaagg ctaaagaagg tatattgctg ttgacagtga gcgnnnnnnn nnnnnnnnnn
     1561 nnnnntagtg aagccacaga tgtannnnnn nnnnnnnnnn nnnnnntgcc tactgcctcg
     1621 gacttcaagg ggctacttta ggagcaatta tcttgtttac taaaactgaa taccttgcta
     1681 tctctttgat acatttttac aaagctgaat taaaatggta taaattaaat cactttacgc
     1741 gtgatctaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     1801 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     1861 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     1921 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     1981 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     2041 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     2101 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     2161 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     2221 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagccaata tgggatcggc
     2281 cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2341 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2401 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     2461 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2521 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2581 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2641 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2701 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2761 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     2821 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     2881 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     2941 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3001 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3061 cgagttcttc tgagcaattc cgcccccccc ccctaacgtt actggccgaa gccgcttgga
     3121 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     3181 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3241 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3301 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3361 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3421 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     3481 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     3541 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaacg tctaggcccc
     3601 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     3661 ggcccagtcc aagcacggcc tgaccaagga gatgaccatg aagtaccgca tggagggctg
     3721 cgtggacggc cacaagttcg tgatcaccgg cgagggcatc ggctacccct tcaagggcaa
     3781 gcaggccatc aacctgtgcg tggtggaggg cggccccttg cccttcgccg aggacatctt
     3841 gtccgccgcc ttcatgtacg gcaaccgcgt gttcaccgag tacccccagg acatcgtcga
     3901 ctacttcaag aactcctgcc ccgccggcta cacctgggac cgctccttcc tgttcgagga
     3961 cggcgccgtg tgcatctgca acgccgacat caccgtgagc gtggaggaga actgcatgta
     4021 ccacgagtcc aagttctacg gcgtgaactt ccccgccgac ggccccgtga tgaagaagat
     4081 gaccgacaac tgggagccct cctgcgagaa gatcatcccc gtgcccaagc agggcatctt
     4141 gaagggcgac gtgagcatgt acctgctgct gaaggacggt ggccgcttgc gctgccagtt
     4201 cgacaccgtg tacaaggcca agtccgtgcc ccgcaagatg cccgactggc acttcatcca
     4261 gcacaagctg acccgcgagg accgcagcga cgccaagaac cagaagtggc acctgaccga
     4321 gcacgccatc gcctccggct ccgccttgcc ctgaagatca ttaattaaga attatcaagc
     4381 ttatcgatac cgtcgacctg cagccaagct tatcgataaa ataaaagatt ttatttagtc
     4441 tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa
     4501 cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca gatcaaggtt
     4561 aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     4621 cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca gcagtttcta
     4681 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     4741 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     4801 taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg cgtcgcccgg
     4861 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     4921 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atgggtaaca
     4981 gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa gtaatcctga
     5041 ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag ttcattatgg
     5101 acagcgcaga aagagctggg gagaattgtg aaattgttat ccgctcacaa ttccacacaa
     5161 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5221 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5281 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5341 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5401 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5461 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5521 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5581 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5641 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5701 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5761 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5821 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5881 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5941 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6001 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6061 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6121 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6181 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6241 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6301 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6361 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6421 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6481 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6541 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6601 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6661 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6721 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6781 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6841 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6901 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6961 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7021 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7081 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7141 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7201 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7261 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     7321 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     7381 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     7441 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     7501 actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     7561 catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc
     7621 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt
     7681 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa tggtgcatgc
     7741 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     7801 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     7861 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     7921 aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact
     7981 caacaatatc accagctgaa gcctatagag tacgagccat agataaaata aaagatttta
     8041 tttagtctcc agaaaaaggg gggaa
//
