LOCUS       sequence-489762-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In-Vitro Synthesis of mRNA encoding a Honeybee-optimized 
            mEmerald-labeled GAP43 Membrane Anchor (MEME) Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4733)
  TITLE     In-Vitro Synthesis of mRNA for Fluorescent Labeling of Honeybee 
            Embryos
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4733)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBSII-MEME-mEmerald(AM)
FEATURES             Location/Qualifiers
     source          1..4733
                     /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    1656..1790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1693..1712)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1747..1766
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2545..2565)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2545..2563)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2584..2600)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2584..2600)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2597..2619)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2608..2624
                     /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(2632..2638,2639..2656,2657..2662))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2677..2698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2815..2832)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2986..3574)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3066..3085)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3745..4536,4537..4605))
                     /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     4368..4387
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4606..4710)
                     /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 aattgttcac cggcgtggtg ccaattttgg tggaattgga cggcgacgtg aacggccaca
      901 aattctcggt gtcgggcgaa ggcgaaggcg acgccaccta cggcaaattg accttgaaat
      961 tcatttgtac caccggcaaa ttgccagtgc catggccaac cttggtgacc accttgacct
     1021 acggcgtgca atgtttcgcc agatacccag accacatgaa acaacacgac ttcttcaaat
     1081 cggccatgcc agaaggctac gtgcaagaaa gaaccatttt cttcaaagac gacggcaact
     1141 acaaaaccag agccgaagtg aaattcgaag gcgacacctt ggtgaacagg attgaattga
     1201 aaggcattga cttcaaagaa gatggcaaca ttttgggcca caaattggaa tacaactaca
     1261 actcgcacaa agtgtacatt accgccgaca aacaaaagaa cggcattaaa gtgaacttca
     1321 aaaccagaca caacattgaa gatggctcgg tgcaattggc cgaccactac caacaaaaca
     1381 ccccaattgg cgacggccca gtgttgttgc cagacaacca ctacttgtcg acccaatcga
     1441 aattgtcgaa agacccaaac gaaaagaggg accacatggt gttgttggaa ttcgtgaccg
     1501 ccgccggcat taccttgggc atggacgaat tgtacaaata acctgcaggg atcataatca
     1561 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     1621 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     1681 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     1741 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cccaggaagc
     1801 tcctctgtgt cctcataaac cctaacctcc tctacttgag aggacattcc aatcataggc
     1861 tgcccatcca ccctctgtgt cctcctgtta attaggtcac ttaacaaaaa ggaaattggg
     1921 taggggtttt tcacagaccg ctttctaagg gtaattttaa aatatctggg aagtcccttc
     1981 cactgctgtg ttccagaagt gttggtaaac agcccacaaa tgtcaacagc agaaacatac
     2041 aagctgtcag ctttgcacaa gggcccaaca ccctgctcat caagaagcac tgtggttgct
     2101 gtgttagtaa tgtgcaaaac aggaggcaca ttttccccac ctgtgtaggt tccaaaatat
     2161 ctagtgtttt catttttact tggatcagga acccagcact ccactggata agcattatcc
     2221 ttatccaaaa cagccttgtg gtcagtgttc atctgctgac tgtcaactgt agcatttttt
     2281 ggggttacag tttgagcagg atatttggtc ctgtagtttg ctaacacacc ctgcagctcc
     2341 aaaggttccc caccaacagc aaaaaaatga aaatttgacc cttgaatggg ttttccagca
     2401 ccattttcat gagttttttg tgtccctgaa tgcaagttta acatagcagt taccccaata
     2461 acctcagttt taacagtaac agcttcccac atcaaaatat ttccacaggt taagtcctca
     2521 tttaaagagc tccagctttt gttcccttta gtgagggtta attgcgcgct tggcgtaatc
     2581 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     2641 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     2701 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     2761 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2821 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2881 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2941 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3001 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3061 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3121 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3181 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3241 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3301 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3361 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3421 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3481 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3541 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3601 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3661 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3721 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3781 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3841 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3901 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3961 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4021 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4081 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4141 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4201 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4261 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4321 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4381 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4441 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4501 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4561 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4621 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4681 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cac
//
