LOCUS       sequence-175673-        6569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6569)
  AUTHORS   Morel C, Carlson SM, White FM, Davis RJ
  TITLE     Mcl-1 integrates the opposing actions of signaling pathways that 
            mediate survival and apoptosis.
  JOURNAL   Mol Cell Biol. 2009 Jul . 29(14):3845-52.
  PUBMED    19433446
REFERENCE   2  (bases 1 to 6569)
  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: "Mol Cell 
            Biol. 2009 Jul . 29(14):3845-52."
COMMENT     Sequence Label: pCDNA3.1-hMcl-1
FEATURES             Location/Qualifiers
     source          1..6569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        123..502
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        503..706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     657..677
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     751..770
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        751..769
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1955..1996
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2006..2023
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2046..2063)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2052..2276
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2322..2750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2409..2428)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2619..2640
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2759..2779)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2764..3093
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2944..3079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3006..3025
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3160..3954
                     /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(3214..3233)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3824..3843
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4130..4251
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4167..4186)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4221..4240
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4300..4316)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4300..4316)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4313..4335)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4324..4340
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4348..4354,4355..4372,4373..4378))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4393..4414
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4531..4548)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4702..5290)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4782..4801)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5461..6252,6253..6321))
                     /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     6084..6103
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6322..6426)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6501..6520)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
       61 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      121 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      181 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      241 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      301 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      361 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      421 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      481 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      541 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      601 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      661 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
      721 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
      781 tggctagtta agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattgccct
      841 tatgtttggc ctcaaaagaa acgcggtaat cggactcaac ctctactgtg ggggggccgg
      901 cttgggggcc ggcagcggcg gcgccacccg cccgggaggg cgacttttgg ctacggagaa
      961 ggaggcctcg gcccggcgag agataggggg aggggaggcc ggcgcggtga ttggcggaag
     1021 cgccggcgca agccccccgt ccaccctcac gccagactcc cggagggtcg cgcggccgcc
     1081 gcccattggc gccgaggtcc ccgacgtcac cgcgaccccc gcgaggctgc ttttcttcgc
     1141 gcccacccgc cgcgcggcgc cgcttgagga gatggaagcc ccggccgctg acgccatcat
     1201 gtcgcccgaa gaggagctgg acgggtacga gccggagcct ctcgggaagc ggccggctgt
     1261 cctgccgctg ctggagttgg tcggggaatc tggtaataac accagtacgg acgggtcact
     1321 accctcgacg ccgccgccag cagaggagga ggacgagttg taccggcagt cgctggagat
     1381 tatctctcgg taccttcggg agcaggccac cggcgccaag gacacaaagc caatgggcag
     1441 gtctggggcc accagcagga aggcgctgga gaccttacga cgggttgggg atggcgtgca
     1501 gcgcaaccac gagacggcct tccaaggcat gcttcggaaa ctggacatca aaaacgaaga
     1561 cgatgtgaaa tcgttgtctc gagtgatgat ccatgttttc agcgacggcg taacaaactg
     1621 gggcaggatt gtgactctca tttcttttgg tgcctttgtg gctaaacact tgaagaccat
     1681 aaaccaagaa agctgcatcg aaccattagc agaaagtatc acagacgttc tcgtaaggac
     1741 aaaacgggac tggctagtta aacaaagagg ctgggatggg tttgtggagt tcttccatgt
     1801 agaggaccta gaaggtggca tcaggaatgt gctgctggct tttgcaggtg ttgctggagt
     1861 aggagctggt ttggcatatc taataagata gggcaattct gcagatatcc agcacagtgg
     1921 cggccgctcg agtctagagg gcccgcggtt cgaaggtaag cctatcccta accctctcct
     1981 cggtctcgat tctacgcgta ccggtcatca tcaccatcac cattgagttt aaacccgctg
     2041 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2101 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2161 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2221 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2281 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2341 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2401 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2461 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2521 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2581 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2641 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2701 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     2761 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2821 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2941 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3001 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     3061 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     3121 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     3181 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     3241 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     3301 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     3361 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     3421 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     3481 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     3541 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     3601 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     3661 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     3721 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3781 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3841 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3901 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3961 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4021 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4081 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     4141 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     4201 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     4261 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     4321 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4381 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4441 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4501 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4561 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4621 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4681 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4741 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4801 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4861 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4921 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4981 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5041 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5101 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     5161 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5221 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5281 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5341 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5401 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5461 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5521 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5581 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5641 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5701 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5761 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5821 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5881 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5941 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6001 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6061 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6121 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6181 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6241 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6301 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6361 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6421 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     6481 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     6541 ctccctgctt gtgtgttgga ggtcgctga
//
