LOCUS       sequence-425611-        5895 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 5895)
  TITLE     pcDNA-HA-Ubc7
REFERENCE   2  (bases 1 to 5895)
  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"
COMMENT     Sequence Label: pcDNA-HA-UBE2L3
FEATURES             Location/Qualifiers
     source          1..5895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     25..46
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(165..185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        170..499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      350..485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     412..431
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             566..1360
                     /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(620..639)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1230..1249
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1534..1655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1571..1590)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1625..1644
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1704..1720)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1704..1720)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1717..1739)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1728..1744
                     /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(1752..1758,1759..1776,1777..1782))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1797..1818
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1935..1952)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2106..2694)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2186..2205)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2865..3656,3657..3725))
                     /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     3488..3507
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3726..3830)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3905..3924)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4096..4475
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4476..4679
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4630..4650
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4724..4743
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4724..4742
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4784..4810
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(5309..5327)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5310..5327)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5347..5364)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5353..5577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(5623..5895,1..156)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5710..5729)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
       61 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
      121 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
      181 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
      241 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
      301 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
      361 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
      421 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
      481 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
      541 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
      601 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
      661 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
      721 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
      781 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
      841 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
      901 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
      961 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     1021 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     1081 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     1141 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     1201 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     1261 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     1321 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     1381 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     1441 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     1501 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     1561 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     1621 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     1681 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1741 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1801 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1861 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1921 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1981 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2041 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2101 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2161 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2221 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     2281 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     2341 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     2401 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     2461 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     2521 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     2581 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2641 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2701 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2761 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2821 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2881 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2941 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3001 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3061 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3121 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3181 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3241 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     3301 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     3361 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     3421 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     3481 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3541 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3601 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3661 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3721 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3781 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3841 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     3901 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     3961 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     4021 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     4081 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     4141 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4201 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4261 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4321 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4381 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4441 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     4501 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     4561 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     4621 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     4681 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     4741 gggagaccca agctagcttg gtaccacgcg taccatggat atctatccat atgacgttcc
     4801 agattatgct gctcagatcg ataagcttat ggcggccagc aggaggctga tgaaggagct
     4861 tgaagaaatc cgcaaatgtg ggatgaaaaa cttccgtaac atccaggttg atgaagctaa
     4921 tttattgact tggcaagggc ttattgttcc tgacaacccc ccatatgata agggagcctt
     4981 cagaatcgaa atcaactttc cagcagagta cccattcaaa ccaccgaaga tcacatttaa
     5041 aacaaagatc tatcacccaa acatcgacga aaaggggcag gtctgtctgc cagtaattag
     5101 tgccgaaaac tggaagccag caaccaaaac cgaccaagta atccagtccc tcatagcact
     5161 ggtgaatgac ccccagcctg agcacccgct tcgggctgac ctagctgaag aatactctaa
     5221 ggaccgtaaa aaattctgta agaatgctga agagtttaca aagaaatatg gggaaaagcg
     5281 acctgtggac taatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     5341 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     5401 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     5461 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     5521 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     5581 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     5641 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5701 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5761 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5821 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5881 tttttcgccc tttga
//
