LOCUS       sequence-46429-e        7884 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 7884)
  TITLE     David Wang unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7884)
  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            "Unpublished"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: pcDKIER
FEATURES             Location/Qualifiers
     source          1..7884
                     /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"
     primer_bind     complement(3309..3326)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3315..3539
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3585..4013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3672..3691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3882..3903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4022..4042)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4027..4356
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4207..4342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4269..4288
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4405..5430
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4405..5430
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5560..5681
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5597..5616)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5651..5670
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5730..5746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5730..5746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5743..5765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5754..5770
                     /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(5778..5784,5785..5802,5803..5808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5823..5844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5961..5978)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6132..6717)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6212..6231)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6888..7679,7680..7748))
                     /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     7511..7530
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7749..7853)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atcccccggg ctgcaggaat tcatggataa
      961 aactttatct agagaagagg caaagcagct aatgcaactt ttatgtttgg acatgagttg
     1021 ctggggcaat ttaccactaa tgagaagaca atacttagta aagtgcaaag aatatcatcc
     1081 agataaagga ggaaatgagg aaagcatgaa gcttcttaat tctttatatt taaaactgca
     1141 agactcagtt agcagtgtac atgatttaaa tgaagaagaa gataatatat ggcaatcttc
     1201 tcaggtatac tgcaaagatt tgtgctgcaa taagtttaga ttagttggtg ctatatatgg
     1261 tgaatactat gaagcatata ttatgaagca gtgggatgta tgcattcatg gctacaatca
     1321 cgaatgccaa tgcattcact gtattttaag caaatatcat aaagaaaagt acaaaattta
     1381 tagaaaacca cctgtgtgga tagagtgcta ctgctataag tgctacaggg agtggttttt
     1441 ttttccaatt tcaatgcaaa catttttctt ttggaaagtg attattttta atactgaaat
     1501 acgagcagtt caaccattat tacggtaagc ataaaataat gtttttaaat aattgtttgt
     1561 gtaatattaa atgtatattt ttgttgtata gatacctaca tacggtacac ctgactggga
     1621 tgagtggtgg tcacagttca acacctactg ggaagaagag ttaagatgta atgaatccat
     1681 gccatcttct ccaaaaagaa gtgcaccaga agaggagcct agctgttctc aggcaacccc
     1741 tcctaagaaa aaacatgcat ttgatgcttc tttagaattt cctaaagagt tgttagagtt
     1801 tgtttcacat gctgtattta gtaataagtg tataacgtgc tttgtagtac atactactag
     1861 agaaaaaggg gaagtacttt ataaaaagtt acttcaaaag tatcaatgta gctttataag
     1921 taagcatgca ttttataata ctgttttaat ctttttctta actcctcaca agcatagagt
     1981 tagtgctatt aataacttct gcaaaggtca ttgtactgtt agctttttat tctgtaaagg
     2041 agttaataac ccttatggtc tgtatagtag gatgtgtaga caaccattta atttgtgtga
     2101 agaaaatatt cctggagggt taaaagaaaa tgaatttaat ccagaagatt tatttgggga
     2161 acctaaggaa ccctcattaa gctggaatca aatagctaac tttgctctgg aatttgacat
     2221 agatgatgta tattacttac tggggtcata tataaggttt gctactaagc ctgaagagtg
     2281 tgaaaaatgc agtaaaaatg atgatgctac ccataaaagg gtacatgtgc aaaatcatga
     2341 aaatgctctg cttttgcaag aaagtaaaag ccaaaaaaat gcttgcacac aagccattga
     2401 tagagttata gcagaaagaa gatataactg tgtaacctta actaggaaaa agttactaac
     2461 taaaaggttt aaaaagcttt ttaatgaaat ggataaaatt gttgttggag aaagaaaaat
     2521 actgctttat atggcttcaa tagcctggta tactggacta aataaaaaaa tagatgaatt
     2581 agttgttaga tttttaaagc taattgtaga caacaagcca aaacacagat attggctttt
     2641 caaaggccca attaacagtg gcaaaaccac attagccact gctttgttaa atttgtgtgg
     2701 tggcaaagcc ttgaacatta acataccttc agaaaaacta ccatttgaac ttggagttgc
     2761 tcttgatcaa tatatggttg tgtttgaaga tgtcaaaggt caaataggta ttgaaaagca
     2821 actaccttct ggtaatgggg taaataatct tgataactta agagactacc ttgatggttg
     2881 tgttgaggtt aacttagaaa aaaagcatgt taataagagg tcacaaatat ttcccccagg
     2941 gattgtaacc atgaatgaat attgtattcc agaaacagtt gctgtgagat ttgaaaaaac
     3001 tgttatgttt actattaaaa gaaatttaag agaaagccta gaaaaaacac cacagttact
     3061 atcacaaaga atattacaca gtggaatagc tatgttgtta ttgctaatat ggtatagacc
     3121 tgtaagtgac tttgatgaag aaattcaaag taatgttgtg tactggaaag aagttcttga
     3181 taattatata ggccttactg agtttgccac tatgcaaatg aatgtaacca atggaaaaaa
     3241 catactagaa aagtggtttg aataagtcga cctcgagtct agagggcccg tttaaacccg
     3301 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     3361 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3421 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3481 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     3541 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     3601 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     3661 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     3721 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     3781 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     3841 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     3901 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     3961 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     4021 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4081 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     4141 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     4201 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     4261 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     4321 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     4381 attttcggat ctgatcagca cgtgatgaaa aagcctgaac tcaccgcgac gtctgtcgag
     4441 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     4501 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     4561 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     4621 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     4681 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     4741 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     4801 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     4861 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     4921 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     4981 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     5041 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     5101 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     5161 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     5221 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     5281 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     5341 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     5401 cccagcactc gtccgagggc aaaggaatag cacgtgctac gagatttcga ttccaccgcc
     5461 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     5521 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5581 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5641 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5701 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5761 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5821 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5881 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5941 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6001 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6061 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6121 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6181 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6241 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6301 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6361 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6421 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6481 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6541 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6601 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6661 ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6721 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6781 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6841 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6901 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6961 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7021 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7081 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7141 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7201 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7261 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7321 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7381 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7441 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7501 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7561 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7621 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7681 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7741 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7801 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7861 ccccgaaaag tgccacctga cgtc
//
