LOCUS       sequence-470848-        5651 bp    DNA     circular SYN 01-MAY-2025
DEFINITION  Transfection backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5651)
  TITLE     Promega Early Access Collection
REFERENCE   2  (bases 1 to 5651)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pFC37A HiBiT CMV-neo Flexi Vector
FEATURES             Location/Qualifiers
     source          1..5651
                     /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"
     CDS             1087..1422
                     /codon_start=1
                     /product="ribonuclease from Bacillus amyloliquefaciens"
                     /label=barnase
                     /note="The barnase gene is lethal in standard bacterial 
                     transformation strains."
                     /translation="MAQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLAD
                     VAPGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHY
                     QTFTKIR"
     primer_bind     complement(1597..1615)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    1652..1773
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1663..1682)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1717..1736
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2018..2375
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2018..2209
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2041..2061)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2226..2361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2426..3220
                     /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(2480..2499)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3090..3109
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3284..3332
                     /note="synthetic polyadenylation signal"
     promoter        3488..3592
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3593..3661,3662..4453)
                     /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     complement(3811..3830)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4611..5199)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4691..4710)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5315..5598
                     /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 ggctagcgat cgccatggaa taagtaagga
     1081 atccacatgg cacaggttat caacacgttt gacggggttg cggattatct tcagacatat
     1141 cataagctac ctgataatta cattacaaaa tcagaagcac aagccctcgg ctgggtggca
     1201 tcaaaaggga accttgcaga cgtcgctccg gggaaaagca tcggcggaga catcttctca
     1261 aacagggaag gcaaactccc gggcaaaagc ggacgaacat ggcgtgaagc ggatattaac
     1321 tatacatcag gcttcagaaa ttcagaccgg attctttact caagcgactg gctgatttac
     1381 aaaacaacgg accattatca gacctttaca aaaatcagat aatgtttaat gaccccgtgt
     1441 cgagctcaag gcagttcagg tggctcgagc ggtgtgagcg gctggcggct gttcaagaag
     1501 attagctaag gccgcgactc tagagtcgac ctgcaggcat gcaagctgat ccggctgcta
     1561 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     1621 cccttggggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     1681 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     1741 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     1801 tttcaggttc agggggagat gtgggaggtt tttttaagca agtaaaacct ctacaaatgt
     1861 ggtaaaatcg aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc
     1921 cttacgcatc tgtgcggtat ttcacaccgc atacgcggat ctgcgcagca ccatggcctg
     1981 aaataacctc tgaaagagga acttggttag gtaccttctg aggcggaaag aaccagctgt
     2041 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2101 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2161 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2221 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2281 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2341 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tgattcttct gacacaacag
     2401 tctcgaactt aaggctagag ccaccatgat tgaacaagat ggattgcacg caggttctcc
     2461 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     2521 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     2581 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     2641 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     2701 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     2761 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     2821 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     2881 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     2941 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     3001 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     3061 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     3121 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     3181 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     3241 atgaccgacc aagcgacgcc caacctgcca tcacgatggc cgcaataaaa tatctttatt
     3301 ttcattacat ctgtgtgttg gttttttgtg tgaatcgata gcgataagga tcctctttgc
     3361 gcttgcgttt tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg
     3421 tttctgcgga ctggctttct acgtaatggt ttcttagacg tcaggtggca cttttcgggg
     3481 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3541 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     3601 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3661 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3721 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3781 ttttccaatg atgagcactt tcaaagttct gctatgtggc gcggtattat cccgtattga
     3841 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3901 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3961 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacta tcggaggacc
     4021 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4081 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4141 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4201 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4261 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4321 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4381 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4441 taagcattgg taattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca
     4501 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4561 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4621 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4681 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4741 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4801 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4861 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4921 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4981 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5041 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5101 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5161 tgcaagcagc agattacgcg cagaaaaaaa ggatttcaag aagatccttt gatcttttct
     5221 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5281 tcaaaaagga tcttcaccta gatcctttta tagtccggaa atacaggaac gcacgctgga
     5341 tggcccttcg ctgggatggt gaaaccatga aaaatggcag cttcagtgga ttaagtgggg
     5401 gtaatgtggc ctgtaccctc tggttgcata ggtattcata cggttaaaat ttatcaggcg
     5461 cgattgcggc agtttttcgg gtggtttgtt gccattttta cctgtctgct gccgtgatcg
     5521 cgctgaacgc gttttagcgg tgcgtacaat taagggatta tggtaaatcc acttactgtc
     5581 tgccctcgta gccatcgaga taaaccgcag tactccggcc acgatgcgtc cggcgtagag
     5641 gatcgagatc t
//
