LOCUS       sequence-489761-        4724 bp    DNA     circular SYN 20-OCT-2025
DEFINITION  In-Vitro Synthesis of mRNA encoding a Honeybee-optimized 
            mCherry-labeled GAP43 Membrane Anchor (MEME) Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4724)
  TITLE     In-Vitro Synthesis of mRNA for Fluorescent Labeling of Honeybee 
            Embryos
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4724)
  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            "Unpublished"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-mCherry(AM)
FEATURES             Location/Qualifiers
     source          1..4724
                     /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    1647..1781
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1684..1703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1738..1757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2536..2556)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2536..2554)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2575..2591)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2575..2591)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2588..2610)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2599..2615
                     /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(2623..2629,2630..2647,2648..2653))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2668..2689
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2806..2823)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2977..3565)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3057..3076)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3736..4527,4528..4596))
                     /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     4359..4378
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4597..4701)
                     /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 ggccgcctcg aaaggcgaag
      841 aagataacat ggccattatt aaagaattca tgagattcaa agtgcacatg gaaggctcgg
      901 tgaacggcca cgaattcgaa attgaaggcg aaggcgaagg cagaccatac gaaggcaccc
      961 aaaccgccaa actgaaagtg accaaaggcg gcccactgcc attcgcctgg gacattctgt
     1021 cgccacaatt catgtacggc tcgaaagcct acgtgaaaca cccagccgac attccagact
     1081 acctgaaact gtcgttccca gaaggcttca aatgggaaag agtgatgaac ttcgaagatg
     1141 gcggcgtggt gaccgtgacc caagactcgt cgctgcaaga cggcgaattc atttacaaag
     1201 tgaaactgag aggcaccaac ttcccatcgg acggcccagt gatgcaaaag aaaaccatgg
     1261 gctgggaagc ctcgtcggaa agaatgtacc cagaagatgg cgccctgaaa ggcgaaatta
     1321 aacaaagact gaaactgaaa gacggcggcc actacgacgc cgaagtgaaa accacctaca
     1381 aagccaagaa accagtgcaa ctgccaggcg cctacaacgt gaacattaaa ctggacatta
     1441 cctcgcacaa cgaagattac accattgtgg aacaatacga aagagccgaa ggcagacact
     1501 cgaccggcgg catggacgaa ctgtacaaat aacctgcagg gatcataatc agccatacca
     1561 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     1621 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     1681 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     1741 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccccaggaag ctcctctgtg
     1801 tcctcataaa ccctaacctc ctctacttga gaggacattc caatcatagg ctgcccatcc
     1861 accctctgtg tcctcctgtt aattaggtca cttaacaaaa aggaaattgg gtaggggttt
     1921 ttcacagacc gctttctaag ggtaatttta aaatatctgg gaagtccctt ccactgctgt
     1981 gttccagaag tgttggtaaa cagcccacaa atgtcaacag cagaaacata caagctgtca
     2041 gctttgcaca agggcccaac accctgctca tcaagaagca ctgtggttgc tgtgttagta
     2101 atgtgcaaaa caggaggcac attttcccca cctgtgtagg ttccaaaata tctagtgttt
     2161 tcatttttac ttggatcagg aacccagcac tccactggat aagcattatc cttatccaaa
     2221 acagccttgt ggtcagtgtt catctgctga ctgtcaactg tagcattttt tggggttaca
     2281 gtttgagcag gatatttggt cctgtagttt gctaacacac cctgcagctc caaaggttcc
     2341 ccaccaacag caaaaaaatg aaaatttgac ccttgaatgg gttttccagc accattttca
     2401 tgagtttttt gtgtccctga atgcaagttt aacatagcag ttaccccaat aacctcagtt
     2461 ttaacagtaa cagcttccca catcaaaata tttccacagg ttaagtcctc atttaaagag
     2521 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2581 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2641 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     2701 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     2761 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2821 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2881 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2941 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3001 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3061 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3121 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3181 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3241 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3301 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3361 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3421 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3481 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3541 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3601 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3661 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3721 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3781 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3841 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3901 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3961 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4021 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4081 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4141 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4201 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4261 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4321 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4381 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4441 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4501 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4561 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4621 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4681 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccac
//
