LOCUS       sequence-478490-        6602 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Express HiBiT-ZER1 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 6602)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6602)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: HiBiT-ZER1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..6602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3545..3563)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    3600..3721
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3611..3630)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3665..3684
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3966..4323
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3966..4157
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3989..4009)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4174..4309
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4338..4385
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4417..5211
                     /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(4471..4490)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5081..5100
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5275..5323
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(5562..6150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5642..5661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6266..6549
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgct caccatggtg agcggctggc
     1081 ggctgttcaa gaagattagc gggagctccg gtggctcgag cggtgcgatc gccatggcgt
     1141 ccgacactcc cgagtcgctg atggccctct gtactgactt ctgcttgcgc aacctggatg
     1201 gcaccctggg ctacctgctg gacaaggaga ccctgcggct acatccggac atcttcttgc
     1261 ccagcgagat ctgtgaccgg ctcgtcaatg agtatgtgga gctggtgaac gctgcctgta
     1321 acttcgagcc acacgagagc ttcttcagcc tcttttcgga cccccgcagc acccgcctca
     1381 cgcggatcca cctccgtgag gacctggtgc aggaccagga cctggaggcc atccgcaagc
     1441 aggacctggt ggagctgtac ctgactaact gcgagaagct gtccgccaag agcctgcaga
     1501 cactgaggag cttcagccac accctggtgt ccttgagcct cttcggctgt acaaacattt
     1561 tctatgagga ggagaaccca gggggctgtg aagatgagta cctcgtcaac cccacctgcc
     1621 aggtgctggt taaggatttc accttcgagg gcttcagccg cctccgcttc ctcaacttgg
     1681 gccgcatgat tgattgggtc cctgtggagt ccctgctgcg gccgcttaac tccctggctg
     1741 ccttggacct ctcaggcatt cagacgagcg acgcagcctt cctcacccag tggaaagaca
     1801 gcctggtgtc cctcgtcctc tacaacatgg acctgtccga cgaccacatc cgggtcatcg
     1861 tgcagctgca caagctgcga cacctggaca tctcccgaga ccgcctctcc agctactaca
     1921 agttcaagct gactcgggag gtgctgagcc tctttgtgca gaagctgggg aacctaatgt
     1981 ccctggacat ctctggccac atgatcctag agaactgcag catctccaag atggaagagg
     2041 aagcggggca gaccagcatt gagccttcca agagcagcat catacctttc cgggctctga
     2101 agaggccgct gcagttcctc gggctctttg agaactctct gtgccgcctc acgcacattc
     2161 cagcctacaa agtaagtggt gacaaaaacg aagagcaggt gctgaatgcc atcgaggcct
     2221 acacggagca ccggcctgag atcacctcgc gggccatcaa cttgcttttt gacatcgccc
     2281 gcatcgagcg ttgcaaccag ctgctgcggg ccctgaagct ggtcatcacg gccctcaagt
     2341 gccacaaata tgacaggaac attcaagtga caggcagcgc cgctctcttc tacctaacaa
     2401 attccgagta ccgctcagag cagagtgtga agctgcgccg gcaggttatc caggtggtgc
     2461 tgaatggcat ggaatcctac caggaggtga cggtgcagcg gaactgctgc ctgacgctct
     2521 gcaacttcag catccccgag gagctggaat tccagtaccg ccgggtcaac gagctcctgc
     2581 tcagcatcct caaccccacg cggcaggacg agtctatcca gcggatcgcc gtgcacctgt
     2641 gcaatgccct ggtctgccag gtagacaacg accacaagga ggccgtgggc aagatgggct
     2701 ttgtcgtgac catgctgaag ctgattcaga agaagctgct ggacaagaca tgtgaccagg
     2761 tcatggagtt ctcctggagt gccctgtgga acatcacaga tgaaactcct gacaactgcg
     2821 agatgttcct caatttcaac ggcatgaagc tcttcctgga ctgcctgaag gaattcccag
     2881 agaagcagga actgcatagg aatatgctag gacttttggg gaatgtggca gaagtgaagg
     2941 agctgaggcc tcaactaatg acttcccagt tcatcagcgt cttcagcaac ctgttggaga
     3001 gcaaggccga tgggatcgag gtttcctaca atgcctgcgg cgtcctctcc cacatcatgt
     3061 ttgatggacc cgaggcctgg ggcgtctgtg agccccagcg tgaggaggtg gaggaacgca
     3121 tgtgggctgc catccagagc tgggacataa actctcggag aaacatcaat tacaggtcat
     3181 ttgaaccaat tctccgcctc cttccccagg gaatctctcc tgtcagccag cactgggcaa
     3241 cctgggccct gtataacctc gtgtctgtct acccggacaa gtactgccct ctgctgatca
     3301 aagaaggggg gatgcccctt ctgagggaca taattaagat ggcgaccgca cggcaggaga
     3361 ccaaggaaat ggcccgcaag gtgattgagc actgcagtaa ctttaaagag gagaacatgg
     3421 acacgtctag agtttaaacg aattcgggct cggtacccgg ggatcctcta gagtcgacct
     3481 gcaggcatgc aagctgatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
     3541 gccaccgctg agcaataact agcataaccc cttggggcgg ccgcttcgag cagacatgat
     3601 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     3661 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     3721 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     3781 tttaagcaag taaaacctct acaaatgtgg taaaatcgaa ttttaacaaa atattaacgc
     3841 ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     3901 acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac ttggttaggt
     3961 accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4021 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4081 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4141 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4201 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4261 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4321 aaaaagctta attaactgtt gacaattaat catcggcata gtatatcggc atagtataat
     4381 acgacaaggt gaggaactaa acccaggagg cagatcatga ttgaacaaga tggattgcac
     4441 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4501 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4561 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4621 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4681 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4741 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4801 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4861 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4921 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     4981 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5041 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5101 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5161 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5221 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgatgg ccgcaataaa
     5281 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agcgataagg
     5341 atcctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg
     5401 ggtcagcacc gtttctgcgg actggctttc tacccggtat cagctcactc aaaggcggta
     5461 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5521 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5581 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5641 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5701 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5761 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5821 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5881 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5941 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6001 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6061 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6121 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     6181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6241 atcttcacct agatcctttt atagtccgga aatacaggaa cgcacgctgg atggcccttc
     6301 gctgggatgg tgaaaccatg aaaaatggca gcttcagtgg attaagtggg ggtaatgtgg
     6361 cctgtaccct ctggttgcat aggtattcat acggttaaaa tttatcaggc gcgattgcgg
     6421 cagtttttcg ggtggtttgt tgccattttt acctgtctgc tgccgtgatc gcgctgaacg
     6481 cgttttagcg gtgcgtacaa ttaagggatt atggtaaatc cacttactgt ctgccctcgt
     6541 agccatcgag ataaaccgca gtactccggc cacgatgcgt ccggcgtaga ggatcgagat
     6601 ct
//
