LOCUS       sequence-381088-        7369 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 7369)
  AUTHORS   Liu F, Wagner S, Campbell RB, Nickerson JA, Schiffer CA, Ross AH
  TITLE     PTEN enters the nucleus by diffusion.
  JOURNAL   J Cell Biochem. 2005 Oct 1. 96(2):221-34.
  PUBMED    16088943
REFERENCE   2  (bases 1 to 7369)
  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: "J Cell 
            Biochem. 2005 Oct 1. 96(2):221-34."
COMMENT     Sequence Label: GFP-PTEN
FEATURES             Location/Qualifiers
     source          1..7369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(113..132)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        304..683
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        684..887
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     838..858
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     932..951
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        932..950
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(993..995,996..998,999..1709)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1038..1059)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1299..1318)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1646..1667
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2991..3009)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2992..3009)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3029..3046)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3035..3259
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3305..3733
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3392..3411)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3602..3623
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3742..3762)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3747..4076
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3927..4062
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3989..4008
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4143..4937
                     /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(4197..4216)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4807..4826
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5111..5232
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5148..5167)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5202..5221
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5281..5297)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5281..5297)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5294..5316)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5305..5321
                     /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(5329..5335,5336..5353,5354..5359))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5374..5395
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5512..5529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5683..6271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5763..5782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6442..7233,7234..7302))
                     /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     7065..7084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7303..7369,1..38))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
       61 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
      121 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
      181 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
      241 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc
      301 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
      361 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      421 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      481 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      541 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      601 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      661 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      721 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      781 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      841 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      901 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag
      961 cttgcatgcc tgcaggtcga ctctagactg ccatggtgag caagggcgag gagctgttca
     1021 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1081 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1141 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcgtgc
     1201 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1261 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1321 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1381 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1441 acgtctatat cacggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     1501 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1561 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     1621 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1681 tcactctcgg catggacgag ctgtacaaga aaggatccat gacagccatc atcaaagaga
     1741 tcgttagcag aaacaaaagg agatatcaag aggatggatt cgacttagac ttgacctata
     1801 tttatccaaa cattattgct atgggatttc ctgcagaaag acttgaaggc gtatacagga
     1861 acaatattga tgatgtagta aggtttttgg attcaaagca taaaaaccat tacaagatat
     1921 acaatctttg tgctgaaaga cattatgaca ccgccaaatt taattgcaga gttgcacaat
     1981 atccttttga agaccataac ccaccacagc tagaacttat caaacccttt tgtgaagatc
     2041 ttgaccaatg gctaagtgaa gatgacaatc atgttgcagc aattcactgt aaagctggaa
     2101 agggacgaac tggtgtaatg atatgtgcat atttattaca tcggggcaaa tttttaaagg
     2161 cacaagaggc cctagatttc tatggggaag taaggaccag agacaaaaag ggagtaacta
     2221 ttcccagtca gaggcgctat gtgtattatt atagctacct gttaaagaat catctggatt
     2281 atagaccagt ggcactgttg tttcacaaga tgatgtttga aactattcca atgttcagtg
     2341 gcggaacttg caatcctcag tttgtggtct gccagctaaa ggtgaagata tattcctcca
     2401 attcaggacc cacacgacgg gaagacaagt tcatgtactt tgagttccct cagccgttac
     2461 ctgtgtgtgg tgatatcaaa gtagagttct tccacaaaca gaacaagatg ctaaaaaagg
     2521 acaaaatgtt tcacttttgg gtaaatacat tcttcatacc aggaccagag gaaacctcag
     2581 aaaaagtaga aaatggaagt ctatgtgatc aagaaatcga tagcatttgc agtatagagc
     2641 gtgcagataa tgacaaggaa tatctagtac ttactttaac aaaaaatgat cttgacaaag
     2701 caaataaaga caaagccaac cgatactttt ctccaaattt taaggtgaag ctgtacttca
     2761 caaaaacagt agaggagccg tcaaatccag aggctagcag ttcaacttct gtaacaccag
     2821 atgttagtga caatgaacct gatcattata gatattctga caccactgac tctgatccag
     2881 agaatgaacc ttttgatgaa gatcagcata cacaaattac aaaagtctga gaattctgca
     2941 gatatccatc acactggcgg ccgctcgagc atgcatctag agggccctat tctatagtgt
     3001 cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3061 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3121 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3181 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3241 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     3301 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     3361 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     3421 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     3481 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     3541 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     3601 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     3661 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     3721 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     3781 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3901 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3961 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4021 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4081 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4141 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4201 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4261 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4321 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4381 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4441 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4501 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4561 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4621 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     4681 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4741 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4801 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4861 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4921 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4981 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5041 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5101 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5161 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5221 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5281 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5341 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5401 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5461 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5521 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5581 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5641 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5701 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5761 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5821 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5881 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5941 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6001 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6061 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6121 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6181 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6241 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6301 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6361 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6421 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6481 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6541 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6601 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6661 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6721 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6781 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6841 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6901 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6961 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7021 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7081 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7141 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7201 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7261 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     7321 ttattgaagc atttatcagg gttattgtct catgagcgga tacatattt
//
