LOCUS       40924_10166        8300 bp DNA     circular SYN 15-NOV-2021
DEFINITION  Expression of SLNCR1 lncRNA tagged with 12 copies of the MS2 
            stem-loop sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8300)
  AUTHORS   Schmidt K, Joyce CE, Buquicchio F, Brown A, Ritz J, Distel RJ, Yoon 
            CH, Novina CD
  TITLE     The lncRNA SLNCR1 Mediates Melanoma Invasion through a Conserved 
            SRA1-like Region.
  JOURNAL   Cell Rep. 2016 May 31;15(9):2025-37. doi: 
            10.1016/j.celrep.2016.04.018. Epub 2016 May 19.
  PUBMED    27210747
REFERENCE   2  (bases 1 to 8300)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8300)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.celrep.2016.04.018"; journalName: "Cell Rep"; date: 
            "2016-05-31- 31"; volume: "15"; issue: "9"; pages: "2025-37"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        3162..3180
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3201..3219
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3271..3289
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3310..3328
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3380..3398
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3419..3437
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3489..3507
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3528..3546
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3598..3616
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3637..3655
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3707..3725
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     misc_RNA        3746..3764
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat
                     protein"
     promoter        complement(3853..3871)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    3897..4121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4167..4595
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4609..4938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5005..5796
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5973..6106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6143..6159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6167..6183)
                     /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(6191..6221)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6236..6257)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(6374..6391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6545..7133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7307..8164)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8165..8269)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttcccggg
      901 atacttagga tccagggcgc gcaggcggcg cgggtgcgcg gtgcggcgct ggtatccaga
      961 ggacgcggtc accgcctctg gcatttgtcg ttctgcgctt ctccgcaagg accctctgtt
     1021 aggcaggcgc ccaccgtaag cctcccgggc cttgtgaacc tgcaaaccca agtctgagag
     1081 acgatccgcc ttcagcgctt tccagcttgg cagagaggct ttcccggcgg ggatctttgg
     1141 ttggcgctgg cgatgcgcgg ggaagaaagg cgaggagcgg cgtccaggct gggtgatgtc
     1201 ccagcacgag taggcgggat gcgctcgctt ggtcctccgg gcgcccggtc cctgcccgcg
     1261 tcgcgcgccc acccctgggg acgagaaggc ggccgcctga ggacccccgc ccgcgacctc
     1321 cgcgagtctg gagcgcagag gacagggtct ggctgctctt tggccttgga tggaaagtgg
     1381 ggaattgggt ggggggctgc ggacccctta acgtggatta cttggtgtgt atcagctggg
     1441 ctcagaagac ccacgacctc ttctccatcc gtggattgat ttgttctgct taacagctgg
     1501 gtcgccaagc tggaggtatt tttccctctc caccctggtc ttctcctgta acgtgtggcc
     1561 gccttttcca gcacggcctc ctgccttcct ggtgcacttt ttggagaacg tggtggaatc
     1621 agaggtttct ggctgactcg gtgggtgctt tgaaccagga aaggacaaga aagaggatgg
     1681 gaaggactga tccacattcc caccaggaag tttagcagaa cccccgcgtg ccacctggac
     1741 cccttggaag gacctggctc aggctggacc acctcttgag aggcaggagc tctggatttg
     1801 atcaagaatt ctttgctgag catggtgcct catgcctata atcccaacac tttgggaggc
     1861 cagtgtggga ggatctcttg agcccaggag ttcaagacta gcctgggcaa cacagagaga
     1921 ccccatctct aaaataataa taataataaa ataaaaaatt agcagggcat ggtggcatgt
     1981 gcctgtagtc ccagctaccc aggaggctga ggcaagagga tggctggagc ctgggatgtt
     2041 gaggctgcaa tgaactgtga ttaccccact gcactccagc ctgggcaaaa gagcgagaga
     2101 ccctgtctca aataataata ataataataa tcttattttg gagaataaag agacctctgg
     2161 atttgaggtg ccatttgggt agaaagaaaa gacgtttaca ccgagaaata gtctgtgttg
     2221 ccctgaagga gcagagggat gcatcgctgg aggtgaccta cagttgaaga agactcatta
     2281 tgacagacct tgtccttctt ccttgtggaa agtgtttcct ctgctgctac tgctcatgag
     2341 actcttcccc ctccctgtcc cagggaacca aagggctttc taccacaccc tttcttgccc
     2401 cccgcctccc atgtctgctg tgcctttgta ctcagcaatt cttgtttgct ccattatctt
     2461 ccagccggat acagagtgaa tagttaacca cacttaggtc aaataggatc taaatttttg
     2521 ttcctgctcc gtgtaaagag gccagtgttt gtgtgttgca agcagccttg gaatagtaac
     2581 tcttctcatt tgtttgggat ctggccacca agttccagaa tgatacacgg atcagtgcag
     2641 aagttcatca ggctctcgga ccttagggct gttggagaag gcttcagcag cagaactgat
     2701 ggtgaaggct cgtgttctcc atcctcaact ttctttgctt cgatcataca caagaataca
     2761 tttggaaggg caaaaaatga acactgtcgt tcattgcagc cgtgttttgt gacacagatg
     2821 cacagtctgc tgtgaagacc ttctctcaag tggcatttgg gagtccatgc cagatcatgg
     2881 tgcttcatga gagactgaca gctatcaggg gttgtggcac ttagtgagga ctctcctccc
     2941 ccagtgtgtg ctgatgacac atacacacct gacaatagct tgagtcttct ctgttccttt
     3001 tactcttcta gataagtatc acgtgaagct tggtaccgag ctcggatcca ctagtaacgg
     3061 ccgccagtgt gctggaattc gaatggccat gggacgtcga cctgaggtaa ttataacccg
     3121 ggccctatat atggatccta aggtacctaa ttgcctagaa aacatgagga tcacccatgt
     3181 ctgcaggtcg actctagaaa acatgaggat cacccatgtc tgcagtattc ccgggttcat
     3241 tagatcctaa ggtacctaat tgcctagaaa acatgaggat cacccatgtc tgcaggtcga
     3301 ctctagaaaa catgaggatc acccatgtct gcagtattcc cgggttcatt agatcctaag
     3361 gtacctaatt gcctagaaaa catgaggatc acccatgtct gcaggtcgac tccagaaaac
     3421 atgaggatca cccatgtctg cagtattccc gggttcatta gatcctaagg tacctaattg
     3481 cctagaaaac atgaggatca cccatgtctg caggtcgact ctagaaaaca tgaggatcac
     3541 ccatgtctgc agtattcccg ggttcattag atcctaaggt acctaattgc ctagaaaaca
     3601 tgaggatcac ccatgtctgc aggtcgactc tagaaaacat gaggatcacc catgtctgca
     3661 gtattcccgg gttcattaga tcctaaggta cctaattgcc tagaaaacat gaggatcacc
     3721 catgtctgca ggtcgactcc agaaaacatg aggatcaccc atgtctgcag tattcccggg
     3781 ttcattagat ctgcgcgcga tcgatatcca tcacactggc ggccgctcga gcatgcatct
     3841 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     3901 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3961 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     4021 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     4081 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     4141 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     4201 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4261 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4321 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4381 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4441 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4501 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4561 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     4621 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     4681 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4741 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     4801 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     4861 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     4921 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     4981 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     5041 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     5101 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     5161 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     5221 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     5281 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     5341 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     5401 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     5461 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     5521 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     5581 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     5641 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     5701 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     5761 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     5821 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     5881 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     5941 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     6001 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6061 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     6121 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     6181 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     6241 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     6301 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     6361 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     6421 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     6481 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     6541 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6601 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6661 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6721 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6781 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6841 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6901 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6961 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     7021 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     7081 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     7141 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     7201 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     7261 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     7321 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     7381 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     7441 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     7501 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     7561 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     7621 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     7681 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7741 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7801 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7861 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7921 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7981 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     8041 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     8101 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     8161 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     8221 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     8281 gaaaagtgcc acctgacgtc
//
