LOCUS       sequence-474709-        6472 bp    DNA     circular SYN 05-JUN-2025
DEFINITION  Express LgBiT-CALM1/SmBiT-ADCY1.2 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 6472)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 6472)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: LgBiT-CALM1/SmBiT-ADCY1.2 BiBiT Vector
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    15..96
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        175..532
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        175..366
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(198..218)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      383..518
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      557..566
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             563..961
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    985..1033
                     /note="synthetic polyadenylation signal"
     rep_origin      1159..1747
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1648..1667
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1905..2699)
                     /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(2016..2035)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2626..2645
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2731..2778)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     protein_bind    2788..2821
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    2936..3057
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     3027..3046
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4289..4308)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4290..4308)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4350..4369)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          4352..4484
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        complement(4620..4823)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4649..4669)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4824..5127
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5247..5265
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5247..5264
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             5835..6284
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPT
                     EAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
ORIGIN
        1 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
       61 tcacaaattt cacaaataaa gcattttttt cactgccgac gtcttaatta acgcgtcttg
      121 cgcagcacca tggcctgaaa taacctctga aagaggaact tggttaggta ccttctgagg
      181 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
      241 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
      301 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
      361 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
      421 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
      481 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctcga
      541 ttcttctgac actagcgcca ccatggccaa gcctttgtct caagaagaat ccaccctcat
      601 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
      661 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
      721 gggaccttgt gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
      781 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
      841 ccgacaggtg cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
      901 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
      961 aactctagcc atcacgatgg ccgcaataaa atatctttat tttcattaca tctgtgtgtt
     1021 ggttttttgt gtgacgattc cggactataa aaggatctag gtgaagatcc tttttgataa
     1081 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1141 aaagatcaaa ggatcttctt gaaatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1201 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1261 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     1321 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1381 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1441 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1501 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1561 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1621 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1681 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1741 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1801 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     1861 gtgagctgat accggttggg cgtcgcttgg tcggtcattt cgaatcagaa gaactcgtca
     1921 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
     1981 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
     2041 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
     2101 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
     2161 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
     2221 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
     2281 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
     2341 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     2401 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     2461 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     2521 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     2581 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     2641 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     2701 atctgcctcc tgggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc
     2761 cgatgattaa ttgtcaacag tggatccata acttcgtata atgtatgcta tacgaagtta
     2821 tattagaatt cttaattaag attttaccac atttgtagag gttttacttg cttaaaaaaa
     2881 cctcccacat ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     2941 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3001 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     3061 tgtctgctcg aagcggcctt gctcagcggt ggcagcagcc aaatcagctt gcatgcctgc
     3121 aggtcgactc tagaggatcc ccgggtaccg agcccgaatt cgtttaaact cacgtgcaca
     3181 gtcgtccccg ttgcagggtg ggagaccaaa accccaggag gacatccagg ccactcagtg
     3241 agggatggat ctgaaccttg tcatcgtcct cgcaggtcat gacattggag aaggggatct
     3301 cggccttgaa cattttccgg cagtgcatga gctgcaccag caggtagcag acagtcttga
     3361 acttcatcct tttggccggt tttatatctg agagaatcag gcctggaaat atctttcggc
     3421 gatgggatgg cacaataaaa aaggtttcga tgttatgagt tttcaagaaa ctgttcctct
     3481 catgtccgta acccggttct acctcgtagt ccccattcaa gcacgctagg gtcgtctttg
     3541 tgatatgaac cttccctggc aggccagcgg cttccatgac attggccaag gtcacatcat
     3601 tggaccacac gtcgtactgc cacttgcgca agcccaggac accacagagg accctgcccg
     3661 tgtgcagacc cacacgcatg ttcagatcca cctcggtggc ttcagccaca gatgtgatgg
     3721 tatcaatcat gtcgagtccc atctccacac agcagtgggc atggtcagtc ttgggctggg
     3781 tgaggcccga cacgcagtag tagcagtccc cgagaatctt gatgcggcga cagtggttct
     3841 ccgtggctaa ttcatcgaac ttgccgaaga gctcattgag gagtttcacc agctcctggg
     3901 ctgtgcactg ggatgccaag cccgtgaaac ccacgatgtc agcaaacagg atgctcacat
     3961 tgtcgtgcct ctggatgtaa atcttgtgga aaatcctctc agggggcttc aggaagtcct
     4021 ccttcatctc catggcaacg ttccggggca ggaggctcat gaggagccgc tcctgcttct
     4081 cgttctcatc ctccagcctc agtcggtcct caatgcagct ccgggcctgc aggaacgcct
     4141 tcctctgtga acgctcagtc agaatccgca caaagacccc atacatggcg atcgcaccgc
     4201 tcgagcctcc acctccgctc ccgccaccac cggagctccc gagaatctcc tcgaacagcc
     4261 ggtagccggt caccatggtg agcgctagcc ctatagtgag tcgtattaat actctagcct
     4321 taagagctgt aattgaactg ggagtggaca cctgtggaga gaaaggcaaa gtggatgtca
     4381 gtaagaccaa taggtgccta tcagaaacgc aagagtcttc tctgtctcga caagcccagt
     4441 ttctattggt ctccttaaac ctgtcttgta accttgatac ttacctgccc agtgcctcac
     4501 gaccaacttc tgcagcttaa gttcgagact gttgtgtcag aagcactgac tgcgttagca
     4561 atttaactgt gataaactac cgcaataaag cttctagtga tctgacggtt cactaaacca
     4621 gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt caacggggcg
     4681 gggttattac gacattttgg aaagtcccgt tgattttggt gccaaaacaa actcccattg
     4741 acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc acgcccattg
     4801 gtgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta gatgtactgc
     4861 caagtaggaa agtcccgtaa ggtcatgtac tgggcataat gccaggcggg ccatttaccg
     4921 tcattgacgt caataggggg cggacttggc atatgataca cttgatgtac tgccaagtgg
     4981 gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat tggcgttact
     5041 atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg tcagccaggc
     5101 gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatg aactaatgac
     5161 cccgtaattg attactatta ataactagtg gcgcgccagg gaggcctata taagcagagc
     5221 tggtttagtg aaccgtcaga tcgcccattt aggtgacact atagaataca agctacttgt
     5281 tctttttgca gctagcgctc accatggtct tcacactcga agatttcgtt ggggactggg
     5341 aacagacagc cgcctacaac ctggaccaag tccttgaaca gggaggtgtg tccagtttgc
     5401 tgcagaatct cgccgtgtcc gtaactccga tccaaaggat tgtccggagc ggtgaaaatg
     5461 ccctgaagat cgacatccat gtcatcatcc cgtatgaagg tctgagcgcc gaccaaatgg
     5521 cccagatcga agaggtgttt aaggtggtgt accctgtgga tgatcatcac tttaaggtga
     5581 tcctgcccta tggcacactg gtaatcgacg gggttacgcc gaacatgctg aactatttcg
     5641 gacggccgta tgaaggcatc gccgtgttcg acggcaaaaa gatcactgta acagggaccc
     5701 tgtggaacgg caacaaaatt atcgacgagc gcctgatcac ccccgacggc tccatgctgt
     5761 tccgagtaac catcaacagt gggagctccg gtggtggcgg gagcggaggt ggaggctcga
     5821 gcggtgcgat cgccatggct gatcagctga ccgaagaaca gattgctgaa ttcaaggaag
     5881 ccttctccct atttgataaa gatggcgatg gcaccatcac aacaaaggaa cttggaactg
     5941 tcatgaggtc actgggtcag aacccaacag aagctgaatt gcaggatatg atcaatgaag
     6001 tggatgctga tggtaatggc accattgact tccccgaatt tttgactatg atggctagaa
     6061 aaatgaaaga tacagatagt gaagaagaaa tccgtgaggc attccgagtc tttgacaagg
     6121 atggcaatgg ttatatcagt gcagcagaac tacgtcacgt catgacaaac ttaggagaaa
     6181 aactaacaga tgaagaagta gatgaaatga tcagagaagc agatattgat ggagacggac
     6241 aagtcaacta tgaagaattc gtacagatga tgactgcaaa atgagtttaa acgaattcgg
     6301 gctcggtacc cggggatcct ctagaaacac ctaggctctc gtacgaagtc agctgataac
     6361 gtcgacgtat tgcggccgcg gatcataatc agccatacca catttgtaga ggttttactt
     6421 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tg
//
