LOCUS       sequence-439377-        6646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged Psma4-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6646)
  TITLE     Ori lab plasmids
REFERENCE   2  (bases 1 to 6646)
  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: PSMA4_GFP_Cterm
FEATURES             Location/Qualifiers
     source          1..6646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(12..31)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      825..1413
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1314..1333
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1567..1584
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1701..1722
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1737..1742,1743..1760,1761..1767)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1775..1791
                     /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)."
     primer_bind     1780..1802
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1799..1815
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1799..1815
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1864..1985
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1875..1894)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1929..1948
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(2115..3137)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             complement(2115..3137)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     protein_bind    complement(3145..3192)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     3324..3343
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3476..3700
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3689..3706
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3750..4463,4464..4466,4467..4469))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     4144..4163
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4403..4424
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(4482..4502)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5411..5435)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    5445..5463
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5466..5484
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        complement(5486..5689)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5515..5535)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5690..6069
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6241..6260
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6335..6439
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6440..6508,6509..6646,1..654)
                     /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"
ORIGIN
        1 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
       61 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      121 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      181 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      241 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      301 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      361 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      421 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      481 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      541 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
      601 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
      661 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
      721 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
      781 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
      841 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      901 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      961 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1021 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1081 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1141 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1201 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1261 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1321 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1381 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1441 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1501 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1561 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     1621 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     1681 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     1741 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     1801 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacgg tatacagaca
     1861 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1921 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     1981 aagttggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     2041 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     2101 tctcgtagta cgtgctattc ctttgccctc ggacgagtgc tggggcgtcg gtttccacta
     2161 tcggcgagta cttctacaca gccatcggtc cagacggccg cgcttctgcg ggcgatttgt
     2221 gtacgcccga cagtcccggc tccggatcgg acgattgcgt cgcatcgacc ctgcgcccaa
     2281 gctgcatcat cgaaattgcc gtcaaccaag ctctgataga gttggtcaag accaatgcgg
     2341 agcatatacg cccggagccg cggcgatcct gcaagctccg gatgcctccg ctcgaagtag
     2401 cgcgtctgct gctccataca agccaaccac ggcctccaga agaagatgtt ggcgacctcg
     2461 tattgggaat ccccgaacat cgcctcgctc cagtcaatga ccgctgttat gcggccattg
     2521 tccgtcagga cattgttgga gccgaaatcc gcgtgcacga ggtgccggac ttcggggcag
     2581 tcctcggccc aaagcatcag ctcatcgaga gcctgcgcga cggacgcact gacggtgtcg
     2641 tccatcacag tttgccagtg atacacatgg ggatcagcaa tcgcgcatat gaaatcacgc
     2701 catgtagtgt attgaccgat tccttgcggt ccgaatgggc cgaacccgct cgtctggcta
     2761 agatcggccg cagcgatcgc atccatggcc tccgcgaccg gctgcagaac agcgggcagt
     2821 tcggtttcag gcaggtcttg caacgtgaca ccctgtgcac ggcgggagat gcaataggtc
     2881 aggctctcgc tgaattcccc aatgtcaagc acttccggaa tcgggagcgc ggccgatgca
     2941 aagtgccgat aaacataacg atctttgtag aaaccatcgg cgcagctatt tacccgcagg
     3001 acatatccac gccctcctac atcgaagctg aaagcacgag attcttcgcc ctccgagagc
     3061 tgcatcaggt cggagacgct gtcgaacttt tcgatcagaa acttctcgac agacgtcgcg
     3121 gtgagttcag gctttttggc caaggaagtt cctatacttt ctagagaata ggaacttcgg
     3181 aataggaact tctaggtacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     3241 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     3301 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     3361 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     3421 cagggcgcgt ggggataccc cctagagccc cagctggttc tttccgcctc agaagccata
     3481 gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc
     3541 tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga tgcaatttcc
     3601 tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg
     3661 gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga tcagcgggtt
     3721 taaacgggcc ctctagactc gagtcactat tacttgtaca gctcgtccat gccgagagtg
     3781 atcccggcgg cggtcacgaa ctccttaagg accatgtgat cgcgcttctc gttggggtct
     3841 ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     3901 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc ctcgatgttg
     3961 tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat gatatagacg
     4021 ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     4081 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     4141 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     4201 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     4261 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     4321 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     4381 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     4441 gtgaacagct cctcgccctt gctcaccatg gtggtacctt caccttgaaa atataaattt
     4501 tcgcggccgc tagggcggcc gcggtattta tccttttctt tctgttcttt ttctttcttc
     4561 tcacgctcag ctttggcttc ttcttcctcg tgttttttga tcaactgctc cacttctttt
     4621 tgtttgagaa ctctgattac tgtctttcca ttctctcttg ttagtgttgc aatttccact
     4681 ttttcagcag agagtttact aacatccatg gtcttattta gtactttgat agctaaagca
     4741 agtgctgact tcaaggtcat ttctccttct ttatagtctt gtttcaacat tgacacagct
     4801 gcagcgctat tatttccaat gcatgtggcc ttccatcccc cgtaatttcc actagggtca
     4861 ctctgataga gctgaaagcc atagtgctta tcccagccaa tgtacagcaa tgaaacacca
     4921 aagggacgtt ttcctccaaa ttgtgtataa gcttgtttga tatcacacag cgctgtaacc
     4981 aactgctcac aaggtattgg ctcctgatac tgtaataaat acctttgagc aatgagcctt
     5041 agttcattag tcagaacatt agcatcagaa gttatgcctg ccacactgca agccatgtcc
     5101 tcattgagtt tataaatttt ttcagaaaaa aagacttcat caagaagctt gtggatgttg
     5161 cgtctctctg ctgcaagcaa aacaccatca tttgctaaaa ttcccaaaca ggtgcctgca
     5221 tgtccaatag cttccatggc atattcaact tggtataagc gaccttctgg agaaaatata
     5281 gtggtcctgg agtcatatct tcgagacatg caggcctcct aagcttaagt ttaaacgcta
     5341 gagtccggag gctggatcgg tcccggtgtc ttctatggag gtcaaaacag cgtggatggc
     5401 gtctccaggc gatctgacgg ttcactaaac gagctcgtcg acgatctcta tcactgatag
     5461 ggagatctct atcactgata gggagagctc tgcttatata gacctcccac cgtacacgcc
     5521 taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag tcccgttgat
     5581 tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga aatccccgtg
     5641 agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc atggtaatag
     5701 cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg
     5761 cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat
     5821 gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc
     5881 aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc
     5941 gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact
     6001 ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa ctagtcaata
     6061 atcaatgtca acgcgtatat ctggcccgta catcgcgaag cagcgcaaaa cgcctaaccc
     6121 taagcagatt cttcatgcaa ttgtcggtca agccttgcct tgttgtagct taaattttgc
     6181 tcgcgcacta ctcagcgacc tccaacacac aagcagggag cagatactgg cttaactatg
     6241 cggcatcaga gcagattgta ctgagagtgc accatagggg atcgggagat ctcccgatcc
     6301 gtcgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     6361 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     6421 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     6481 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     6541 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     6601 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcact
//
