LOCUS       sequence-489768-        4730 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  In-Vitro Synthesis of mRNA encoding a Honeybee-optimized 
            mTagBFP2-labeled GAP43 Membrane Anchor (MEME) Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4730)
  TITLE     In-Vitro Synthesis of mRNA for Fluorescent Labeling of Honeybee 
            Embryos
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4730)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 20, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pBSII-MEME-mTagBFP(AM)
FEATURES             Location/Qualifiers
     source          1..4730
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    1653..1787
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1690..1709)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1744..1763
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2542..2562)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2542..2560)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2581..2597)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2581..2597)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2594..2616)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2605..2621
                     /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)."
     promoter        complement(join(2629..2635,2636..2653,2654..2659))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2674..2695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2812..2829)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2983..3571)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3063..3082)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3742..4533,4534..4602))
                     /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     4365..4384
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4603..4707)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcctgc agcccgggca
      721 aaatgttgtg ctgcatgagg aggaccaaac aagtggaaaa gaatgatgag gaccaaaaaa
      781 ttggctcggc cggctcggcc gccggctcgg gcgaattcgc ggccgccatg tcgaaaggtg
      841 aagaattgat taaagaaaac atgcatatga aattgtacat ggaaggcacc gtggacaacc
      901 accacttcaa atgcacctcg gaaggcgaag gcaaaccata cgaaggcacc caaacaatga
      961 gaattaaagt ggtggaaggt ggcccattgc cattcgcctt cgacattttg gccacctcgt
     1021 tcttgtacgg atcgaaaacc ttcattaacc acacccaagg cattccagat ttcttcaaac
     1081 aatcgttccc agaaggcttc acctgggaaa gagtgacaac ctacgaagac ggaggagtgt
     1141 tgaccgccac ccaagacacc tcgttgcaag acggatgctt gatttacaac gtgaaaatta
     1201 gaggagtgaa cttcacatcg aatggcccag tgatgcaaaa gaaaaccttg ggttgggaag
     1261 ccttcaccga aaccttgtac ccagccgatg gtggtttgga aggcagaaac gacatggcgt
     1321 tgaaattggt gggcggctcg cacttgattg ccaacgccaa aaccacctac agatcgaaaa
     1381 aaccggccaa aaacttgaaa atgccaggtg tgtactacgt ggactacaga ttggaaagaa
     1441 ttaaagaagc caacaacgaa acctacgtgg aacaacatga agtggcggtg gcgagatact
     1501 gcgacttgcc gtcgaaattg ggccacaaat tgaactaacc tgcagggatc ataatcagcc
     1561 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1621 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1681 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     1741 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccc aggaagctcc
     1801 tctgtgtcct cataaaccct aacctcctct acttgagagg acattccaat cataggctgc
     1861 ccatccaccc tctgtgtcct cctgttaatt aggtcactta acaaaaagga aattgggtag
     1921 gggtttttca cagaccgctt tctaagggta attttaaaat atctgggaag tcccttccac
     1981 tgctgtgttc cagaagtgtt ggtaaacagc ccacaaatgt caacagcaga aacatacaag
     2041 ctgtcagctt tgcacaaggg cccaacaccc tgctcatcaa gaagcactgt ggttgctgtg
     2101 ttagtaatgt gcaaaacagg aggcacattt tccccacctg tgtaggttcc aaaatatcta
     2161 gtgttttcat ttttacttgg atcaggaacc cagcactcca ctggataagc attatcctta
     2221 tccaaaacag ccttgtggtc agtgttcatc tgctgactgt caactgtagc attttttggg
     2281 gttacagttt gagcaggata tttggtcctg tagtttgcta acacaccctg cagctccaaa
     2341 ggttccccac caacagcaaa aaaatgaaaa tttgaccctt gaatgggttt tccagcacca
     2401 ttttcatgag ttttttgtgt ccctgaatgc aagtttaaca tagcagttac cccaataacc
     2461 tcagttttaa cagtaacagc ttcccacatc aaaatatttc cacaggttaa gtcctcattt
     2521 aaagagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     2581 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2641 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2701 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2761 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2821 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2881 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2941 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3001 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3061 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3121 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3181 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3241 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3301 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3361 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3421 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3481 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3541 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3601 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3661 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3721 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3781 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3841 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3901 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3961 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     4021 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     4081 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     4141 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     4201 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     4261 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     4321 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     4381 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     4441 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     4501 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     4561 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4621 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4681 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
//
