LOCUS       sequence-475095-        7235 bp    DNA     circular SYN 24-JUN-2025
DEFINITION  Express HiBiT-GABBR1 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 7235)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 7235)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: HiBiT-GABBR1 Fusion Vector
FEATURES             Location/Qualifiers
     source          1..7235
                     /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(4178..4196)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4233..4354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4244..4263)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4298..4317
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4599..4956
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4599..4790
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4622..4642)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4807..4942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4971..5018
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5050..5844
                     /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(5104..5123)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5714..5733
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5908..5956
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(6195..6783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6275..6294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6899..7182
                     /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 caccatgaac tccttctcca
     1081 caagcgcctt cggtccagtt gccttctccc tgggcctgct cctggtgttg cctgctgcct
     1141 tccctgcccc agtctccgtg agcggctggc ggctgttcaa gaagattagc gggagctccg
     1201 gtggctcgag cggtgcgatc gccccgggcg cgggcggggc gcagaccccc aacgccacct
     1261 cagaaggttg ccagatcata cacccgccct gggaaggggg catcaggtac cggggcctga
     1321 ctcgggacca ggtgaaggct atcaacttcc tgccagtgga ctatgagatt gagtatgtgt
     1381 gccgggggga gcgcgaggtg gtggggccca aggtccgcaa gtgcctggcc aacggctcct
     1441 ggacagatat ggacacaccc agccgctgtg tccgaatctg ctccaagtct tatttgaccc
     1501 tggaaaatgg gaaggttttc ctgacgggtg gggacctccc agctctggac ggagcccggg
     1561 tggatttccg gtgtgacccc gacttccatc tggtgggcag ctcccggagc atctgtagtc
     1621 agggccagtg gagcaccccc aagccccact gccaggtgaa tcgaacgcca cactcagaac
     1681 ggcgcgcagt gtacatcggg gcactgtttc ccatgagcgg gggctggcca gggggccagg
     1741 cctgccagcc cgcggtggag atggcgctgg aggacgtgaa tagccgcagg gacatcctgc
     1801 cggactatga gctcaagctc atccaccacg acagcaagtg tgatccaggc caagccacca
     1861 agtacctata tgagctgctc tacaacgacc ctatcaagat catccttatg cctggctgca
     1921 gctctgtctc cacgctggtg gctgaggctg ctaggatgtg gaacctcatt gtgctttcct
     1981 atggctccag ctcaccagcc ctgtcaaacc ggcagcgttt ccccactttc ttccgaacgc
     2041 acccatcagc cacactccac aaccctaccc gcgtgaaact ctttgaaaag tggggctgga
     2101 agaagattgc taccatccag cagaccactg aggtcttcac ttcgactctg gacgacctgg
     2161 aggaacgagt gaaggaggct ggaattgaga ttactttccg ccagagtttc ttctcagatc
     2221 cagctgtgcc cgtcaaaaac ctgaagcgcc aggatgcccg aatcatcgtg ggacttttct
     2281 atgagactga agcccggaaa gttttttgtg aggtgtacaa ggagcgtctc tttgggaaga
     2341 agtacgtctg gttcctcatt gggtggtatg ctgacaattg gttcaagatc tacgaccctt
     2401 ctatcaactg cacagtggat gagatgactg aggcggtgga gggccacatc acaactgaga
     2461 ttgtcatgct gaatcctgcc aatacccgca gcatttccaa catgacatcc caggaatttg
     2521 tggagaaact aaccaagcga ctgaaaagac accctgagga gacaggaggc ttccaggagg
     2581 caccgctggc ctatgatgcc atctgggcct tggcactggc cctgaacaag acatctggag
     2641 gaggcggccg ttctggtgtg cgcctggagg acttcaacta caacaaccag accattaccg
     2701 accaaatcta ccgggcaatg aactcttcgt cctttgaggg tgtctctggc catgtggtgt
     2761 ttgatgccag cggctctcgg atggcatgga cgcttatcga gcagcttcag ggtggcagct
     2821 acaagaagat tggctactat gacagcacca aggatgatct ttcctggtcc aaaacagata
     2881 aatggattgg agggtccccc ccagctgacc agaccctggt catcaagaca ttccgcttcc
     2941 tgtcacagaa actctttatc tccgtctcag ttctctccag cctgggcatt gtcctagctg
     3001 ttgtctgtct gtcctttaac atctacaact cacatgtccg ttatatccag aactcacagc
     3061 ccaacctgaa caacctgact gctgtgggct gctcactggc tttagctgct gtcttccccc
     3121 tggggctcga tggttaccac attgggagga accagtttcc tttcgtctgc caggcccgcc
     3181 tctggctcct gggcctgggc tttagtctgg gctacggttc catgttcacc aagatttggt
     3241 gggtccacac ggtcttcaca aagaaggaag aaaagaagga gtggaggaag actctggaac
     3301 cctggaagct gtatgccaca gtgggcctgc tggtgggcat ggatgtcctc actctcgcca
     3361 tctggcagat cgtggaccct ctgcaccgga ccattgagac atttgccaag gaggaaccta
     3421 aggaagatat tgacgtctct attctgcccc agctggagca ttgcagctcc aggaagatga
     3481 atacatggct tggcattttc tatggttaca aggggctgct gctgctgctg ggaatcttcc
     3541 ttgcttatga gaccaagagt gtgtccactg agaagatcaa tgatcaccgg gctgtgggca
     3601 tggctatcta caatgtggca gtcctgtgcc tcatcactgc tcctgtcacc atgattctgt
     3661 ccagccagca ggatgcagcc tttgcctttg cctctcttgc catagttttc tcctcctata
     3721 tcactcttgt tgtgctcttt gtgcccaaga tgcgcaggct gatcacccga ggggaatggc
     3781 agtcggaggc gcaggacacc atgaagacag ggtcatcgac caacaacaac gaggaggaga
     3841 agtcccggct gttggagaag gagaaccgtg aactggaaaa gatcattgct gagaaagagg
     3901 agcgtgtctc tgaactgcgc catcaactcc agtctcggca gcagctccgc tcccggcgcc
     3961 acccaccgac acccccagaa ccctctgggg gcctgcccag gggaccccct gagccccccg
     4021 accggcttag ctgtgatggg agtcgagtgc atttgcttta taaggtttaa acgaattcgg
     4081 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagctga tccggctgct
     4141 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     4201 ccccttgggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     4261 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     4321 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4381 gtttcaggtt cagggggaga tgtgggaggt ttttttaagc aagtaaaacc tctacaaatg
     4441 tggtaaaatc gaattttaac aaaatattaa cgcttacaat ttcctgatgc ggtattttct
     4501 ccttacgcat ctgtgcggta tttcacaccg catacgcgga tctgcgcagc accatggcct
     4561 gaaataacct ctgaaagagg aacttggtta ggtaccttct gaggcggaaa gaaccagctg
     4621 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4681 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4741 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4801 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4861 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     4921 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc ttaattaact gttgacaatt
     4981 aatcatcggc atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaacccagg
     5041 aggcagatca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     5101 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     5161 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     5221 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     5281 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     5341 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     5401 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     5461 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     5521 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     5581 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5641 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5701 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5761 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5821 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5881 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt
     5941 gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt
     6001 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct
     6061 ttctacccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6121 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6361 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6601 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt
     6781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6841 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttatagtcc
     6901 ggaaatacag gaacgcacgc tggatggccc ttcgctggga tggtgaaacc atgaaaaatg
     6961 gcagcttcag tggattaagt gggggtaatg tggcctgtac cctctggttg cataggtatt
     7021 catacggtta aaatttatca ggcgcgattg cggcagtttt tcgggtggtt tgttgccatt
     7081 tttacctgtc tgctgccgtg atcgcgctga acgcgtttta gcggtgcgta caattaaggg
     7141 attatggtaa atccacttac tgtctgccct cgtagccatc gagataaacc gcagtactcc
     7201 ggccacgatg cgtccggcgt agaggatcga gatct
//
