LOCUS       sequence-474716-        5387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express CA2-NanoLuc(R) Fusion Protein in Mammalian Cells under a CMV
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5387)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5387)
  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            "Unpublished"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: CA2-NanoLuc Fusion Vector
FEATURES             Location/Qualifiers
     source          1..5387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        89..292
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     243..263
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          428..560
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     543..562
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     604..623
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        604..622
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1437..1949
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2037..2055)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2092..2213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2103..2122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2157..2176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2458..2815
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2458..2649
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2481..2501)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2666..2801
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        2830..2877
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2909..3703
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2963..2982)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3573..3592
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3767..3815
                     /note="synthetic polyadenylation signal"
     rep_origin      3972..4517
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4418..4437
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4634..4651
                     /label=L4440
                     /note="L4440 vector, forward primer"
     enhancer        join(5096..5387,1..88)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
       61 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
      121 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      181 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
      241 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      301 gaaccgtcag atcactagaa gctttattgc ggtagtttat cacagttaaa ttgctaacgc
      361 agtcagtgct tctgacacaa cagtctcgaa cttaagctgc agaagttggt cgtgaggcac
      421 tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga aactgggctt
      481 gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta ctgacatcca
      541 ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt aaggctagag
      601 tattaatacg actcactata gggctagcga tcgccatgtc ccatcactgg gggtacggca
      661 aacacaacgg acctgagcac tggcataagg acttccccat tgccaaggga gagcgccagt
      721 cccctgttga catcgacact catacagcca agtatgaccc ttccctgaag cccctgtctg
      781 tttcctatga tcaagcaact tccctgagga tcctcaacaa tggtcatgct ttcaacgtgg
      841 agtttgatga ctctcaggac aaagcagtgc tcaagggagg acccctggat ggcacttaca
      901 gattgattca gtttcacttt cactggggtt cacttgatgg acaaggttca gagcatactg
      961 tggataaaaa gaaatatgct gcagaacttc acttggttca ctggaacacc aaatatgggg
     1021 attttgggaa agctgtgcag caacctgatg gactggccgt tctaggtatt tttttgaagg
     1081 ttggcagcgc taaaccgggc cttcagaaag ttgttgatgt gctggattcc attaaaacaa
     1141 agggcaagag tgctgacttc actaacttcg atcctcgtgg cctccttcct gaatccttgg
     1201 attactggac ctacccaggc tcactgacca cccctcctct tctggaatgt gtgacctgga
     1261 ttgtgctcaa ggaacccatc agcgtcagca gcgagcaggt gttgaaattc cgtaaactta
     1321 acttcaatgg ggagggtgaa cccgaagaac tgatggtgga caactggcgc ccagctcagc
     1381 cactgaagaa caggcaaatc aaagcttcct tcaaagtttc tctcggctcg agcggcgtct
     1441 tcacactcga agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag
     1501 tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc gtaactccga
     1561 tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc
     1621 cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt
     1681 accctgtgga tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg
     1741 gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg
     1801 acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc
     1861 gcctgatcaa ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct
     1921 ggcggctgtg cgaacgcatt ctggcgtaag gccgcgactc tagagtcgac ctgcaggcat
     1981 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2041 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     2101 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2161 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2221 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     2281 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     2341 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     2401 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     2461 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2521 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2581 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2641 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2701 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2761 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2821 taattaactg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     2881 gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc
     2941 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3001 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3061 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3121 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3181 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3241 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3301 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3361 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3421 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3481 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3541 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3601 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3661 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3721 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     3781 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatccgctc
     3841 atgcgatgat aagctgtcaa acatgagaat tacaacttat atcgtatggg gctgacttca
     3901 ggtgctacat ttgaagagat aaattgcact gaaatctaga aatattttat ctgattaata
     3961 agatgatctt cttgagatcg ttttggtctg cgcgtaatct cttgctctga aaacgaaaaa
     4021 accgccttgc agggcggttt ttcgaaggtt ctctgagcta ccaactcttt gaaccgaggt
     4081 aactggcttg gaggagcgca gtcaccaaaa cttgtccttt cagtttagcc ttaaccggcg
     4141 catgacttca agactaactc ctctaaatca attaccagtg gctgctgcca gtggtgcttt
     4201 tgcatgtctt tccgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     4261 ctgaacgggg ggttcgtgca tacagtccag cttggagcga actgcctacc cggaactgag
     4321 tgtcaggcgt ggaatgagac aaacgcggcc ataacagcgg aatgacaccg gtaaaccgaa
     4381 aggcaggaac aggagagcgc acgagggagc cgccaggggg aaacgcctgg tatctttata
     4441 gtcctgtcgg gtttcgccac cactgatttg agcgtcagat ttcgtgatgc ttgtcagggg
     4501 ggcggagcct atggaaaaac ggctttgccg cggccctctc acttccctgt taagtatctt
     4561 cctggcatct tccaggaaat ctccgccccg ttcgtaagcc atttccgctc gccgcagtcg
     4621 aacgaccgag cgtagcgagt cagtgagcga ggaagcggaa tatatcctgt atcacatatt
     4681 ctgctgacgc accggtgcag ccttttttct cctgccacat gaagcacttc actgacaccc
     4741 tcatcagtgc caacatagta agccagtata cactccgcta gcgctgatgt ccggcggtgc
     4801 ttttgccgtt acgcaccacc ccgtcatgag attacgcgtt ttagcggtgc gtacaattaa
     4861 gggattatgg taaatccact tactgtctgc cctcgtagcc atcgagataa accgcagtac
     4921 tccggccacg atgcgtccgg cgtagaggat cgagatcttc aatattggcc attagccata
     4981 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
     5041 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttggcat
     5101 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     5161 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     5221 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     5281 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     5341 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatg
//
