LOCUS       sequence-152190-        5745 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 5745)
  TITLE     Charles Vinson Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5745)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CMV500 A-ATF2
FEATURES             Location/Qualifiers
     source          1..5745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             902..925
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             929..961
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     promoter        complement(1204..1222)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1205..1222)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1242..1259)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1248..1472
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1518..2031
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1605..1624)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1815..1836
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2054..2074)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2059..2388
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2239..2374
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2455..3249
                     /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(2509..2528)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3119..3138
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3423..3557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3460..3479)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3514..3533
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3588..3604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3588..3604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3601..3623)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3612..3628
                     /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(3636..3642,3643..3660,3661..3666))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3681..3702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3819..3836)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3990..4578)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4070..4089)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4749..5540,5541..5609))
                     /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     5372..5391
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5610..5714)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctccaccat
      901 ggactacaag gacgacgatg acaagcatat ggctagcatg actggtggac agcaaatggg
      961 tcgggatcct gacctggaac aacgtgctga ggaactggcc cgtgaaaacg aagagctgga
     1021 aaaagaggcc gaagagctgg agcaggaact ggcagaactc gagaagaaag ctgaagactt
     1081 gagttcatta aatggtcagc tgcagagtga agtcaccctg ctgagaaatg aagtggcaca
     1141 gctgaaacag cttcttctgg ctcataaaga ttgcccttaa ttcaagcttc tagagggccc
     1201 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     1261 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1321 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1381 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1441 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     1501 ctcgaggggg gatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1561 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1621 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1681 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1741 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1801 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1861 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     1921 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaaatatt
     1981 tgcttataca atcttcctgt ttttggggct tttctgatta tcaaccgggg tgggtaccga
     2041 gctcgaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2101 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2161 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2221 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2281 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     2341 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     2401 gcttggatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     2461 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     2521 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     2581 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     2641 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     2701 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     2761 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     2821 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     2881 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     2941 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3001 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3061 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3121 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3181 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3241 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3301 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3361 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     3421 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     3481 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     3541 cttatcatgt ctggatccgt cgacctcgag agcttggcgt aatcatggtc atagctgttt
     3601 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3661 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3721 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3781 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     3841 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     3901 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3961 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4021 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4081 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4141 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4201 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4261 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4321 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4381 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4441 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4501 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4561 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4621 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     4681 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     4741 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4801 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4861 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     4921 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4981 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5041 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5101 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5161 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5221 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5281 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5341 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5401 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5461 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5521 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5581 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5641 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5701 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
