LOCUS       sequence-324068-        7798 bp    DNA     circular SYN 14-APR-2025
DEFINITION  ubb promoter driven expression of lck:mNeonGreen.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7798)
  AUTHORS   Adikes RC, Kohrman AQ, Martinez MAQ, Palmisano NJ, Smith JJ, 
            Medwig-Kinney TN, Min M, Sallee MD, Ahmed OB, Kim N, Liu S, Morabito
            RD, Weeks N, Zhao Q, Zhang W, Feldman JL, Barkoulas M, Pani AM, 
            Spencer SL, Martin BL, Matus DQ
  TITLE     Visualizing the metazoan proliferation-quiescence decision in vivo.
  JOURNAL   Elife. 2020 Dec 22;9. pii: 63265. doi: 10.7554/eLife.63265.
  PUBMED    33350383
REFERENCE   2  (bases 1 to 7798)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2020-12-22"; volume: "9. pii: 63265. doi"; pages: " 
            10.7554/eLife.63265"
COMMENT     Sequence Label: pRM27
FEATURES             Location/Qualifiers
     source          1..7798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..124
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(125..193,194..985)
                     /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(343..362)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1156..1744
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1645..1664
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1898..1915
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2032..2053
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2068..2073,2074..2091,2092..2098)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2106..2122
                     /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     2111..2133
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2130..2146
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2130..2146
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2165..2185
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2167..2185
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             5999..6706
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     regulatory      6254..6263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    6716..6850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6740..6759)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6794..6813
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(7151..7170)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7152..7170)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7180..7197)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7180..7196)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7189..7211)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7338..7765
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7424..7443)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7634..7655
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
       61 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      121 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      181 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      241 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      301 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
      361 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
      421 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
      481 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
      541 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
      601 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
      661 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
      721 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
      781 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
      841 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
      901 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
      961 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1021 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1081 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1141 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1201 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1261 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1321 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1381 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1441 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1501 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1561 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1621 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     1681 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tgaggggggc ggagcctatg
     1741 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     1801 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     1861 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     1921 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     1981 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
     2041 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     2101 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     2161 gcgcgcaatt aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg
     2221 ctctagtgga tctgctgggc ttgctgaagg tagggggtca agaaccagag gtgtaaagta
     2281 cttgagtaat tttacttgat tactgtactt aagtattatt tttggggatt tttactttac
     2341 ttgagtacaa ttaaaaatca atacttttac ttttacttaa ttacattttt ttagaaaaaa
     2401 aagtactttt tactccttac aattttattt acagtcaaaa agtacttatt ttttggagat
     2461 cacttgggcc cggctcgaga ccagcaaagt tctagaattt gtcgaaacat ttatgttata
     2521 tatttcctga aaaaaattct gagtaagttc ttaagtgtta ttgccagcaa cataaacaac
     2581 agacggcaaa atgaataaat gataacaaag cagtaggctt aaataaacct aatttttata
     2641 ggctgttctc tacaaccctc aaacagtgat tagttttgta cttataaact tgccctttca
     2701 ttcatatttc aagaaaattg gttcagaaga tctggatatt ctagcagttg ttcaagctca
     2761 tggagggatc agtgacctga ttccacaatg actaggccta atccagaaat tagatgactg
     2821 tcaacataaa aaggcacagc actcactagc tgccctatat attttattat attttacata
     2881 tattatttta tttatttagc tctgagtgct gtactttctg gttaaagaaa actgcttaca
     2941 acagctaacc tgtactacct caggctcagg gaatttggaa caggtttgtc tggtttgttt
     3001 ctttaaccat gcatgcttgt tttcaactat ggcaacacag tcacatggga cattacagaa
     3061 atgatttgtc gatgacatgc gacttttctt taataaagcg caaagatccc aaaaagcaaa
     3121 cttttaacaa aaatcatata attatatttt caatccagct ttgtagcaac tttgtgctgc
     3181 tgttcactca gcaacagata gtcagtataa ggtcagtgtg tctcaaagca gtgccatctg
     3241 tttcacacat tgcgttctat atataagtgt gctggttgac acgacactgt ataaggccta
     3301 ggctaaaaca caaacaatgt agaatgacac tgtgtttttt ttgtaaacaa atgttgtttt
     3361 tggttaaaca tctttgtgaa aacatcctcc tgtcatgtat ttgctatatt caaatgttaa
     3421 acccgtgcag aatagaacat atacaaaaaa aaacaacaca acacattttt aaacattatt
     3481 aaatatcaag tattgctggc agttctgttt ctgttttaca gtaccctttg ccacagttct
     3541 ccgcttttcc tggtccagat tccacaagtc tgattcacca atagcaaagc gaataaacaa
     3601 ccaaagcagc caatcactgc ttgtagactg tcctgcgaga ccggcccatt ccagcacatt
     3661 ctggaaactt cctttatatg ataattataa atacatttaa attattgata caaaacatgt
     3721 aattcctaga acataaccat agcaatcatt agttttcagg gtaattatgt atttttagga
     3781 tttgactgcg gaaagatctg gtcatgtgac gtctcatgaa cgtcacggcc ctgggtttct
     3841 ataaatacag taggactctc gaccatcggc agatttttcg aagaagaaga tcagtttcag
     3901 gagccgtact gttccgtttc aacgcaaata ttaacggtaa gagcgaattt cctagtttgt
     3961 tttcatgcca ttctttaaaa ccatagcgta ttactttaat tatagtaaac tttcgctttc
     4021 tttattacaa gagacgtttt gtgttgattc tccgcggaca ttttcggtca gacaatcaga
     4081 aaatgaccgc ggaggaccag taacttgcat tacacgtaag ttaaatcttc gtgtattaaa
     4141 atggttaggt tgttaacgtc aaataggtta ccgtgtttgc gtgtgatcag gttggttttg
     4201 ttagattttt gtcagtattt ttaatttatt tgttttagtt tatttatttt ttttgctgaa
     4261 tcatagtttg tgaacaaaga acccggatgt tacatacagt acagccgcca tgttacagag
     4321 agttataact taatcatttt aaaaataatt ttgccttact tttagtttgt catgttgaga
     4381 aatgaggaaa tgttaaaatg aggaaatatc caattaattt aatatatcaa aataatccat
     4441 gattacaatg cactgaactg gagaaaatta agatgttttc tagtgtcatg aaacaaatgt
     4501 aagagatgta cattgtagat gttttatgtc aagaattggc tagttgatgc agcatactgg
     4561 cgatactcag ttgtaataac agtaacgtta catgttaata gactactgag tatgctgttc
     4621 tgtctatgta tgctctgtaa gctacgagaa ggactttttt aaacagtaaa gggtgcaata
     4681 tttttacaaa ttgaattaaa taaaggctgt ctattaagta atatgcttga tatttttctt
     4741 acttgatcgg aaataagaaa aaatataaac gttgttgctc taaaaatcct agttcagttt
     4801 agccaaccac aaataccttt ttgttcctcc aacagttttt ttttcttctc tataatattt
     4861 ggcagtctat agtactccaa atgtttcccc acagtctaac taattggtac agccaaaatc
     4921 atgacactta ttgcaataat aattttggtt cattggcatt gttgatagcc tgtgccacta
     4981 atatggtcga ttgatcatgc ttcaggaaga aaactatatt gtttgatgta agattattaa
     5041 atcttcacct gcttccatta caaactattc ccatcttatt gaattctggt atgtcttaaa
     5101 ggattagttc acttcccaaa tcaaaattta cttagccttt tttcatccat gatccctttt
     5161 tttcatcatt aatgaagaaa ttgtttttga aaaagtttca agattttttt ctctatattg
     5221 tggagcttgt tagtttaaaa ttccaaaatg caatatgtgg cttcaaatgg ttctaaatga
     5281 tcccagtcaa ggaataacag tcttatctaa tgaaaccatt agacctttaa aaataaaaaa
     5341 aataaaagta tttattttta aatgactgag tgattaagtt gaatttcagc gtttccttac
     5401 tgtgtagaag tccttcctta ctggccccac cctttggttc tctgccaatc tgctacctaa
     5461 tgtaatgttg tggaacatta ttattcttta tttcttaatt ttttattttt tattttaaaa
     5521 caatgtaaac tgcacagatg tgcagtttgt ttaaaatggc caatgctttg gaaatgcatg
     5581 acataattag atttcatgat gcacaaagcc aaatctcaga gcttgtgcaa aatgagctat
     5641 catttcacta ggtaagaccc taaattttca tataggatca tttggacaat tttgctgcag
     5701 gtaaaatgca ttctatagtc cactgtcagc cattgttttg gatagtattt atttttctct
     5761 acaagtatag tcaatagttt tctattattt taaaggtttg taacatttaa gggtgaccaa
     5821 atgcaaagta aaatttcatt ttcgggtgaa ctatctcgtt taacatggga gaagtgcaaa
     5881 acatacatta ttggctagaa cattgtagta ttttttaaat ggaaatgtgt gattgctaat
     5941 cttactttga atttgtttac agggatccat gggctgcgtg tgcagcagca accccgagat
     6001 ggtgagcaag ggcgaggagg ataacatggc ctctctccca gcgacacatg agttacacat
     6061 ctttggctcc atcaacggtg tggactttga catggtgggt cagggcaccg gcaatccaaa
     6121 tgatggttat gaggagttaa acctgaagtc caccaagggt gacctccagt tctccccctg
     6181 gattctggtc cctcatatcg ggtatggctt ccatcagtac ctgccctacc ctgacgggat
     6241 gtcgcctttc caggccgcca tggtagatgg ctccggatac caagtccatc gcacaatgca
     6301 gtttgaagat ggtgcctccc ttactgttaa ctaccgctac acctacgagg gaagccacat
     6361 caaaggagag gcccaggtga aggggactgg tttccctgct gacggtcctg tgatgaccaa
     6421 ctcgctgacc gctgcggact ggtgcaggtc gaagaagact taccccaacg acaaaaccat
     6481 catcagtacc tttaagtgga gttacaccac tggaaatggc aagcgctacc ggagcactgc
     6541 gcggaccacc tacacctttg ccaagccaat ggcggctaac tatctgaaga accagccgat
     6601 gtacgtgttc cgtaagacgg agctcaagca ctccaagacc gagctcaact tcaaggagtg
     6661 gcaaaaggcc tttaccgatg tgatgggcat ggacgagctg tacaagatcg atgatgatcc
     6721 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     6781 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     6841 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     6901 ggaggttttt taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctct
     6961 agatcagatc taatactcaa gtacaatttt aatggagtac ttttttactt ttactcaagt
     7021 aagattctag ccagatactt ttacttttaa ttgagtaaaa ttttccctaa gtacttgtac
     7081 tttcacttga gtaaaatttt tgagtacttt ttacacctct gtcaagaacc atatgccggt
     7141 acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg
     7201 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     7261 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     7321 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     7381 tacgcgcagc gtgaccacca cacccgccgc gccctagcgc ccgctccttt cgctttcttc
     7441 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     7501 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     7561 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     7621 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     7681 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     7741 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtgg
//
