LOCUS       sequence-243720-        6472 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 6472)
  AUTHORS   Wu P, Shui W, Carlson BL, Hu N, Rabuka D, Lee J, Bertozzi CR
  TITLE     Site-specific chemical modification of recombinant proteins produced
            in mammalian cells by using the genetically encoded aldehyde tag.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Mar 3. 106(9):3000-5.
  PUBMED    19202059
REFERENCE   2  (bases 1 to 6472)
  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: "Proc Natl 
            Acad Sci U S A. 2009 Mar 3. 106(9):3000-5."
COMMENT     Sequence Label: pcDNA3.1 Neo(+) hSUMF1
FEATURES             Location/Qualifiers
     source          1..6472
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1829..1846)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1835..2059
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2105..2533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2192..2211)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2402..2423
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2542..2562)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2547..2876
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2727..2862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2789..2808
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2943..3737
                     /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(2997..3016)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3607..3626
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3911..4032
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3948..3967)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4002..4021
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4081..4097)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4081..4097)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4094..4116)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4105..4121
                     /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(4129..4135,4136..4153,4154..4159))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4174..4195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4312..4329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4483..5071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4563..4582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5242..6033,6034..6102))
                     /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     5865..5884
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6103..6207)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6282..6301)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcatgg ctgcgcccgc actagggctg gtgtgtggac gttgccctga gctgggtctc
      721 gtcctcttgc tgctgctgct ctcgctgctg tgtggagcgg cagggagcca ggaggccggg
      781 accggtgcgg gcgcggggtc ccttgcgggt tcttgcggct gcggcacgcc ccagcggcct
      841 ggcgcccatg gcagttcggc agccgctcac cgatactcgc gggaggctaa cgctccgggc
      901 cccgtacccg gagagcggca actcgcgcac tcaaagatgg tccccatccc tgctggagta
      961 tttacaatgg gcacagatga tcctcagata aagcaggatg gggaagcacc tgcgaggaga
     1021 gttactattg atgcctttta catggatgcc tatgaagtca gtaatactga atttgagaag
     1081 tttgtgaact caactggcta tttgacagag gctgagaagt ttggcgactc ctttgtcttt
     1141 gaaggcatgt tgagtgagca agtgaagacc aatattcaac aggcagttgc agctgctccc
     1201 tggtggttac ctgtgaaagg cgctaactgg agacacccag aagggcctga ctctactatt
     1261 ctgcacaggc cggatcatcc agttctccat gtgtcctgga atgatgcggt tgcctactgc
     1321 acttgggcag ggaagcggct gcccacggaa gctgagtggg aatacagctg tcgaggaggc
     1381 ctgcataata gacttttccc ctggggcaac aaactgcagc ccaaaggcca gcattatgcc
     1441 aacatttggc agggcgagtt tccggtgacc aacactggtg aggatggctt ccaaggaact
     1501 gcgcctgttg atgccttccc tcccaatggt tatggcttat acaacatagt ggggaacgca
     1561 tgggaatgga cttcagactg gtggactgtt catcattctg ttgaagaaac gcttaaccca
     1621 aaaggtcccc cttctgggaa agaccgagtg aagaaaggtg gatcctacat gtgccatagg
     1681 tcttattgtt acaggtatcg ctgtgctgct cggagccaga acacacctga tagctctgct
     1741 tcgaatctgg gattccgctg tgcagccgac cgcctgccca ctatggactg actcgagtct
     1801 agagggcccg tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     1861 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     1921 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     1981 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     2041 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2101 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2161 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2221 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2281 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     2341 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     2401 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     2461 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2521 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2581 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2641 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2701 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2761 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     2821 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     2881 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     2941 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3001 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3061 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3121 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     3181 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     3241 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     3301 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     3361 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     3421 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     3481 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     3541 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     3601 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     3661 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     3721 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     3781 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3841 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3901 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3961 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4021 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4081 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4141 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4201 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     4261 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     4321 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4381 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4441 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4501 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4561 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4621 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4681 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4741 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4801 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4861 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4921 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4981 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5041 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5101 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5161 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5221 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5281 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5341 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     5401 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5461 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5521 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5581 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5641 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5701 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5761 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5821 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5881 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5941 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6001 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6061 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6121 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6181 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga
     6241 cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc
     6301 gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg
     6361 agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt
     6421 agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg tt
//
