LOCUS       sequence-421232-        7437 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 7437)
  AUTHORS   Galli A, DeFelice LJ, Duke BJ, Moore KR, Blakely RD
  TITLE     Sodium-dependent norepinephrine-induced currents in 
            norepinephrine-transporter-transfected HEK-293 cells blocked by 
            cocaine and antidepressants.
  JOURNAL   J Exp Biol. 1995 Oct . 198(Pt 10):2197-212.
  PUBMED    7500004
REFERENCE   2  (bases 1 to 7437)
  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 Exp Biol.
            1995 Oct . 198(Pt 10):2197-212."
COMMENT     Sequence Label: hNET pcDNA3
FEATURES             Location/Qualifiers
     source          1..7437
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(21..38)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(192..780)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(272..291)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(951..1742,1743..1811))
                     /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     1574..1593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1812..1916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1991..2010)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2182..2561
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2562..2765
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2716..2736
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2810..2829
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2810..2828
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4983..4999)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4983..4999)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5009..5027)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5010..5027)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5047..5064)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5053..5277
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5323..5751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5410..5429)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5620..5641
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5760..5780)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5804..6014
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        5826..6022
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5873..6008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5935..5954
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6089..6883
                     /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(6143..6162)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6753..6772
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7057..7178
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7094..7113)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7148..7167
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7227..7243)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7227..7243)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7240..7262)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7251..7267
                     /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(7275..7281,7282..7299,7300..7305))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7320..7341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
       61 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
      121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
      181 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      241 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      301 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      361 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
      421 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      481 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      541 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      601 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
      661 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      721 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      781 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      841 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      901 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      961 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     1021 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     1081 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     1141 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     1201 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     1261 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     1321 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     1381 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     1441 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     1501 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     1561 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     1621 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     1681 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     1741 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1801 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1861 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1921 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1981 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     2041 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     2101 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     2161 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
     2221 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     2281 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     2341 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2401 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2461 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2521 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     2581 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     2641 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
     2701 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     2761 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
     2821 actataggga gacccaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt
     2881 gctggaattc tgcagatatc catcacactg gcggccgctc gaggtcgagc tacgtcaacg
     2941 gccggacaca gcctcggcgt gcccccagga ccggtaaagt tcctctcgcc agccgcatcc
     3001 atgcttctgg cgcggatgaa cccgcaggtg cagcccgaga acaacggggc ggacacgggt
     3061 ccagagcagc cccttcgggc gcgcaaaact gcggagctgc tggtggtgaa ggagcgcaac
     3121 ggcgtccagt gcctgctggc gccccgcgac ggcgacgcgc agccccggga gacctggggc
     3181 aagaagatcg acttcctgct gtccgtagtc ggcttcgcag tggacctggc caacgtgtgg
     3241 cgcttcccct acctctgcta caagaacggc ggcggtgcct tcttgatccc gtacacactg
     3301 ttccttatca tcgcggggat gcccctgttc tacatggagc tggctctggg acagtacaac
     3361 cgggaggggg ctgccaccgt ttggaaaatc tgcccattct tcaaaggcgt tggctatgct
     3421 gtcatcctga tcgccctgta cgttggcttc tactacaacg tcatcatcgc ctggtcactc
     3481 tactacctct tctcctcctt caccctcaac ctgccctgga ccgactgtgg ccacacctgg
     3541 aacagcccca actgtaccga ccccaagctc ctcaatggct ccgtgcttgg caaccacacc
     3601 aagtactcca agtacaagtt cacgccggca gccgagtttt atgagcgtgg tgtcctgcac
     3661 cttcacgaga gcagcgggat tcatgacatc ggcctgcccc agtggcagct cttgctctgt
     3721 ctgatggtcg tcgtcatcgt cttgtatttt agcctctgga aaggggtgaa gacatcagga
     3781 aaggtggtgt ggatcacagc cacgctgcct tacttcgtgc tgttcgtgct cctggtccat
     3841 ggcgtcacgc tgcccggagc ctccaatggc atcaatgcct acctgcacat cgacttctac
     3901 cgcttgaaag aggccacggt atggattgat gccgcaactc agatattttt ttccttgggg
     3961 gctggatttg gagtattgat tgcatttgcc agttacaaca aatttgacaa caactgttac
     4021 agggatgccc tgctgaccag cagcatcaac tgtatcacca gcttcgtctc tgggttcgcc
     4081 atcttctcca tccttggtta catggcccat gaacacaagg tcaacattga ggatgtggcc
     4141 acagaaggag ctggcctagt gttcatcctg tatccagagg ccatttctac cctgtctgga
     4201 tctacattct gggctgttgt gtttttcgtc atgctcctgg cgctgggcct tgacagctca
     4261 atgggaggca tggaggctgt catcacgggc ctggcagatg acttccaggt cctgaagcga
     4321 caccggaaac tcttcacatt tggcgtcacc ttcagcactt tccttctcgc cctgttctgc
     4381 ataaccaagg gtggaattta cgtcttgacc ctcctggaca cctttgctgc gggcacctcc
     4441 atcctttttg ctgtcctcat ggaagccatc ggagtttcct ggttttatgg agtggacagg
     4501 ttcagcaacg acatccagca gatgatgggg ttcaggccgg gtctatactg gagactgtgc
     4561 tggaagttcg tcagtcctgc cttcctcctg ttcgtggttg tggtcagcat catcaacttc
     4621 aagccactca cctacgacga ctacatcttc ccgccctggg ccaactgggt ggggtggggc
     4681 atcgccctgt cctccatggt cctggtgccc atctacgtca tctataagtt cctcagcacg
     4741 cagggctctc tttgggagag actggcctat ggcatcacgc cagagaacga gcaccacctg
     4801 gtggctcaga gggacatcag acagttccag ttgcaacact ggctggccat ctgagcctgc
     4861 ctggaggaga aggaggaacc cccatgccaa tgtccaggtc acaggcatcc gctgctacgt
     4921 caacgcgttg acgtagctcg acggtatcga taagcttgat atcgaattcc tgcagcccgg
     4981 gggatccact agttctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
     5041 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     5101 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     5161 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     5221 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     5281 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     5341 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5401 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5461 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5521 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5581 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5641 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5701 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     5761 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5821 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     5881 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5941 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     6001 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     6061 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     6121 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     6181 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     6241 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     6301 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     6361 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6421 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6481 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6541 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6601 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6661 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6721 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6781 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6841 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     6901 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     6961 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     7021 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     7081 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7141 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     7201 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     7261 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7321 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7381 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgt
//
