LOCUS       sequence-215206-        7192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of membrane-tagged super folder GFP from zebrafish 
            beta-actin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7192)
  AUTHORS   Wagner DE, Weinreb C, Collins ZM, Briggs JA, Megason SG, Klein AM
  TITLE     Single-cell mapping of gene expression landscapes and lineage in the
            zebrafish embryo.
  JOURNAL   Science. 2018 Apr 26. pii: science.aar4362. doi: 
            10.1126/science.aar4362.
  PUBMED    29700229
REFERENCE   2  (bases 1 to 7192)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2018 Apr 26. pii: science.aar4362. doi: 10.1126/science.aar4362."
COMMENT     Sequence Label: pMTB
FEATURES             Location/Qualifiers
     source          1..7192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      55..64
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      109..118
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(931..950)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(932..950)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    957..1091
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(981..1000)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1035..1054
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(1183..1320,1321..1332)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(1391..1408)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1391..1407)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1400..1422)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1843..1864
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2027..2131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2132..2200,2201..2992)
                     /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(2350..2369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3163..3751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3652..3671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3905..3922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4039..4060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4075..4080,4081..4098,4099..4105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4113..4129
                     /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     4118..4140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4137..4153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4137..4153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4172..4192
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4174..4192
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     4229..4245
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4229..4245
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     misc_feature    join(4279..4290,4291..4478)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
ORIGIN
        1 atttaggtga cactatagaa gagtacaagc tacttgttct ttttgcagga tcccgccacc
       61 atgggctgca tcaagagcaa gcgcaaggac aacctgaacg acgacgaggc cgccatgggc
      121 tgcatcaaga gcaagcgcaa ggacaacctg aacgacgacg agggagcgcc agctggaggt
      181 gcaggtgcag caatgtctaa aggtgaagaa ctgttcaccg gtgttgttcc gatcctggtt
      241 gaactggatg gtgatgttaa cggccacaaa ttctctgttc gtggtgaagg tgaaggtgat
      301 gcaaccaacg gtaaactgac cctgaaattc atctgcacta ccggtaaact gccggttcca
      361 tggccgactc tggtgactac cctgacctat ggtgttcagt gtttttctcg ttacccggat
      421 cacatgaagc agcatgattt cttcaaatct gcaatgccgg aaggttatgt acaggagcgc
      481 accatttctt tcaaagacga tggcacctac aaaacccgtg cagaggttaa atttgaaggt
      541 gatactctgg tgaaccgtat tgaactgaaa ggcattgatt tcaaagagga cggcaacatc
      601 ctgggccaca aactggaata taacttcaac tcccataacg tttacatcac cgcagacaaa
      661 cagaagaacg gtatcaaagc taacttcaaa attcgccata acgttgaaga cggtagcgta
      721 cagctggcgg accactacca gcagaacact ccgatcggtg atggtccggt tctgctgccg
      781 gataaccact acctgtccac ccagtctgtt ctgtccaaag acccgaacga aaagcgcgac
      841 cacatggtgc tgctggagtt cgttactgca gcaggtatca cgcacggcat ggatgagctc
      901 tacaaataag aattcatacg tatccggact ccctatagtg agtcgtatta accggtgatc
      961 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1021 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1081 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1141 gggaggtttt ttgatatccc gggtttaaac gcatgcagat ctaatactca agtacaattt
     1201 taatggagta cttttttact tttactcaag taagattcta gccagatact tttactttta
     1261 attgagtaaa attttcccta agtacttgta ctttcacttg agtaaaattt ttgagtactt
     1321 tttacacctc tgtcaagaac catatgccgg tacccaattc gccctatagg acgtcgtatt
     1381 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     1441 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     1501 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctagc
     1561 gcccgctcct ttcgctttct tcccttcctt tcgcgccgcg tgactggcga atgggacgcg
     1621 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1681 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1741 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     1801 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     1861 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     1921 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     1981 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2041 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2101 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2161 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2221 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2281 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2341 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2401 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2461 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2521 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2581 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2641 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2701 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2761 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2821 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2881 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2941 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3001 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3061 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     3301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     3361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     3421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     3481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3781 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3841 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3901 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3961 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     4021 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     4081 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     4141 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
     4201 gctggagctc caccgcggtg gcggccgctc tagaactagt ggatctgctg ggcttgctga
     4261 aggtaggggg tcaagaacca gaggtgtaaa gtacttgagt aattttactt gattactgta
     4321 cttaagtatt atttttgggg atttttactt tacttgagta caattaaaaa tcaatacttt
     4381 tacttttact taattacatt tttttagaaa aaaaagtact ttttactcct tacaatttta
     4441 tttacagtca aaaagtactt attttttgga gatcacttgg gcccggctcg agggcctagt
     4501 cattacctat actcaacgtt tactctgaga aacttccata ttttagaggt aaaatgctca
     4561 gaattgtaaa aagtcttgtc aaaatacttg gaatttgcca aaaacaaaat aaaagtcaca
     4621 tcacctcatt gtccttggta tcagcaagca cttttaacac aatcagctgt ttttgtttat
     4681 aactcccact gaaggccatg tgcattactc ccaagttaca cagtaaaaag tcacatgtag
     4741 tttatcatga tcactgctgc tcaagcacag ctttatctgt tattcagtgt ggcaatatct
     4801 gaaatgtgtg caaaatatta tttattgtag ctgatcacca acactttcag tgtttttttt
     4861 tttctacttt taatgagaat tatgttctta ggatttatcc ttataaacct ctttaaaaag
     4921 aaatgttatg tctgcataga cacgacatga aattatgaac agaaaattta actatttaag
     4981 atcaagactt gacatataac atttctaatg aacattaaac ctgaggtagg cccacttgca
     5041 aaaatgtaac tgctatcact cactttatac atcattgtgt aacttgtaca cacaaaaata
     5101 gtgaagcaat gtggcatgga atgcaggtcc cattcattca aagttattaa agcactttct
     5161 taaagtgcat tactactgag ctaagaccag ctagttgctc atacaacata ctacaagtag
     5221 tgtcatccta atttggataa aaaaaatgtc cattttatac atggaaaagt actcattaga
     5281 tcccccattg tttgtattac ggtatttcgt gaacacaaga ggtaaatgac ctacagagct
     5341 gctgttgtgt tagattgaaa acacaacaca ggatcatgga ggcatccatc attatgacgc
     5401 attctgccac ttgtaactct gcacagactt tttgaaaagt ttaattgagt cttggcgttg
     5461 tcgcagaaaa atacgaaaaa ttctgtgtta aaatttgcaa ataaaaattc cagttttata
     5521 ggaaatcgcc atggccttgt tctttaacca gttaagcttc cccttctttc actctcaagt
     5581 tgcaagaagc aagtgtagca atgtgcacgc gacagccggg tgtgtgacgc tggaccaatc
     5641 agagcacaga gctccgaaag tttacctttt atggctagag ccgggtatgt gccgtcatat
     5701 aaaagagcgc gcccagcttt tcagcctcac tttgagctcc tccacacgca gctagtgcgg
     5761 aatatcatct gcttgtaacc cattctctta agtcgacaac cccccaaacc caaggtgagt
     5821 tggttttaca gtttttaagc gtttaattat agtttctatc tattaatgtt aagtaaaatg
     5881 tgtaaatgga taaatatgat ttgatgaagt ctttagacag ttcataacgg tctctttgta
     5941 gttatctact aggtttgatt tataagtggt ggttcttttg ataatctatg ttatatttaa
     6001 taaatattgg ctgtatctta acttttttgt agcgttagtg gatgtggcag gtgagaatgt
     6061 cggtaattta acgtgaccag tcgtaggcac gacattgaac attgaatagg ccggtgtgaa
     6121 atgggtgtta agtcttaata taaagttgtg ttctggctaa tctctcttta acagccataa
     6181 atgtgatttt taactacagt tttgagggtg ctggtgagca tgttgcacac ttgatggata
     6241 gccggcatgg gaagttcttt gtgcaggcag tgctgcagca gggtgtgacc tactttagct
     6301 agccggctaa ccagctctca tctgatgtaa cgtaaacccc atcaatctaa catcggctca
     6361 ttcgtgcttt ataagatcga agatcttgtt attagtaacg ggtattggca tttagtttta
     6421 acttggttaa ttttcttttt gttatgtgtt ctatttaaat gtagcctgca gccccctctg
     6481 tctttaatga agctgaagat taacgttaac gttagtgaag tgttgtatcc gctatgaaag
     6541 atttgaattt tagaggtgtg gtccacttta ataacggctg cggagttctc tggctgcact
     6601 tcagccgctg tgactgtcga agcagagccc atcagttttt caggatctcg gtgatttttg
     6661 aggctagact atgaactgaa ccgactgaga gtgaccgtcc agaaatgctt tagtgtgaag
     6721 tctccgctca cgaggagtcc gtggactgac tgcagatctg tgacgcagta acacccggcg
     6781 gcagacaccc gttgaaatgc ggttgtgtaa ttgatcgcag gcgagggtgg aagaggatgt
     6841 gaaacttcat ttgtgtaaaa tttagggagt ggtcctggtg tgatgaatgt cgaaatctgt
     6901 tcctttttac tgagccctac gactctggct gagtgccaca ccgccggcag ccgcaaagcg
     6961 gctcaaacca ttgcctttta tggtaataat gagagaatgc agagggactt cctttgtctg
     7021 gcatatctga ggcgcgcgct gtcactagcg cccaccagcg gtcagactgt agaatgcaga
     7081 gcaaaacagg aagttgactc cagatggtca catgcttgct gaaacgcctc tccctggcag
     7141 cagtgcaatt ctaacagttt ccctttcttt tacagttcgc catcctaggc ct
//
